The arginine-phenylalanine-amide neuropeptide receptor family comprises a subclass within the G protein-coupled receptor superfamily with crucial roles in physiological regulation.These receptors recognize and bind ne...The arginine-phenylalanine-amide neuropeptide receptor family comprises a subclass within the G protein-coupled receptor superfamily with crucial roles in physiological regulation.These receptors recognize and bind neuropeptides with an arginine-phenylalanine-amide motif,thereby participating in a variety of biological processes such as energy metabolism,pain perception,and reproductive functions.In this review,we explore the physiological and pathological processes involving these receptors and delve into the structure-activity relationships of their ligand peptides,clarifying the key structural motifs within these neuropeptides that determine their biological activity,pharmacological potency,and receptor selectivity.Particular emphasis is placed on their roles in modulating nociception,regulating appetite,and maintaining reproductive health.Additionally,we discuss the therapeutic potential of structure-based drug design targeting these receptors based on existing cryo-electron microscopy structures.The available structural insights into ligand-binding pockets and G protein-receptor interaction interfaces provide a clear perspective and valuable complement to ligand optimization.展开更多
From a neuroscience perspective,cancer neuroscience has emerged as a subfield of cancer research.Presumable mechanisms underlying cancer-related neuronal activity(termed neurosciences)include the induction and modulat...From a neuroscience perspective,cancer neuroscience has emerged as a subfield of cancer research.Presumable mechanisms underlying cancer-related neuronal activity(termed neurosciences)include the induction and modulation of signaling pathways that govern cell fate determination and emotional responses(anxiety and stress),such as structural molecules(synaptic structures and current transduction)and secretory substances(neurotransmitters,cytokines,hormones and neuropeptides).In the past 3 years,these neuronal activities,which can either promote cancer growth or be hijacked by cancer cells to support tumor survival and invasion,have been widely demonstrated to be closely related to cancer progression.The molecular mechanisms are also being refined.Despite their great promise,translating neuroscientific discoveries into clinically actionable strategies for cancer diagnosis,prognosis,and treatment remains a formidable task.In this comprehensive review,we attempt to provide a full account of the intersection between neuroscience and cancer research.From the perspective of cancer neuroscience,we fully discuss the potential signaling molecules and their regulatory mechanisms,as well as targets and emerging therapeutic strategies that control tumor progression via multiomics approaches.Overall,cancer neuroscience may have unprecedented potential for understanding neuronal functions and cancer development,ultimately offering the significantly improved cancer treatment.展开更多
Ultraviolet(UV)-induced photoaging skin has become an urgent issue.The functional foods and cosmetics aiming to improve skin photoaging are developing rapidly,and the demand is gradually increasing year by year.Collag...Ultraviolet(UV)-induced photoaging skin has become an urgent issue.The functional foods and cosmetics aiming to improve skin photoaging are developing rapidly,and the demand is gradually increasing year by year.Collagen peptides have been proven to display diverse physiological activities,such as excellent moisture retention activity,hygroscopicity,tyrosinase inhibitory activity and antioxidant activity,which indicates that they have great potential in amelioration of UV-induced photoaging.The main objective of this article is to recap the main mechanisms to improve photoaging skin by collagen peptides and their physiological activities in photo-protection.Furthermore,the extraction and structural characteristics of collagen peptides are overviewed.More importantly,some clinical trials on the beneficial effect on skin of collagen peptides are also discussed.In addition,prospects and challenges of collagen peptides are emphatically elucidated in this review.This article implies that collagen peptides have great potential as an effective ingredient in food and cosmetics industry with a wide application prospect.展开更多
In the present study,we investigate the expression profile of the epidermal growth factor receptor family,which comprises EGFR/ ErbBl,HER2/ErbB2,HER3/ErbB3 and HER4/ErbB4 in oral leukoplakia(LP).The expression of four...In the present study,we investigate the expression profile of the epidermal growth factor receptor family,which comprises EGFR/ ErbBl,HER2/ErbB2,HER3/ErbB3 and HER4/ErbB4 in oral leukoplakia(LP).The expression of four epidermal growth factor receptor (EGFR) family genes and their ligands were measured in LP tissues from 14 patients and compared with levels in 10 patients with oral lichen planus(OLP) and normal oral mucosa(NOM) from 14 healthy donors by real-time polymerase chain reaction(PCR) and immunohistochemistry.Synchronous mRNA coexpression of ErbBl,ErbB2,ErbB3and ErbB4 was detected in LP lesions.Out of the receptors,only ErbB4 mRNA and protein was more highly expressed in LP compared with NOM tissues.These were strongly expressed by epithelial keratinocytes in LP lesions,as shown by immunohistochemistry.Regarding the ligands,the mRNA of Neuregulin2 and 4 were more highly expressed in OLP compared with NOM tissues.Therefore,enhanced ErbB4 on the keratinocytes and synchronous modulation of EGFR family genes may contribute to the pathogenesis and carcinogenesis of LP.展开更多
Ginger(Zingiber officinale),the type species of Zingiberaceae,is one of the most widespread medicinal plants and spices.Here,we report a high-quality,chromosome-scale reference genome of ginger‘Zhugen’,a traditional...Ginger(Zingiber officinale),the type species of Zingiberaceae,is one of the most widespread medicinal plants and spices.Here,we report a high-quality,chromosome-scale reference genome of ginger‘Zhugen’,a traditionally cultivated ginger in Southwest China used as a fresh vegetable,assembled from PacBio long reads,Illumina short reads,and high-throughput chromosome conformation capture(Hi-C)reads.The ginger genome was phased into two haplotypes,haplotype 1(1.53 Gb with a contig N50 of 4.68 M)and haplotype 0(1.51 Gb with a contig N50 of 5.28 M).Homologous ginger chromosomes maintained excellent gene pair collinearity.In 17,226 pairs of allelic genes,11.9%exhibited differential expression between alleles.Based on the results of ginger genome sequencing,transcriptome analysis,and metabolomic analysis,we proposed a backbone biosynthetic pathway of gingerol analogs,which consists of 12 enzymatic gene families,PAL,C4H,4CL,CST,C3’H,C3OMT,CCOMT,CSE,PKS,AOR,DHN,and DHT.These analyses also identified the likely transcription factor networks that regulate the synthesis of gingerol analogs.Overall,this study serves as an excellent resource for further research on ginger biology and breeding,lays a foundation for a better understanding of ginger evolution,and presents an intact biosynthetic pathway for species-specific gingerol biosynthesis.展开更多
The CRISPR-Cas13 system,an RNA-guided editing tool,has emerged as a highly efficient and stable RNA editing technique.Although the CRISPR-Cas13 system has been developed in several insect species,its application in le...The CRISPR-Cas13 system,an RNA-guided editing tool,has emerged as a highly efficient and stable RNA editing technique.Although the CRISPR-Cas13 system has been developed in several insect species,its application in lepidopterans has not yet been reported.In the present study,we evaluated the RNA cleavage activity of the CRISPR-Cas13 system in the silkworm(Bombyx mori),a model lepidopteran insect,both ex vivo and in vivo.We established two stable silkworm BmE cell lines expressing PspCas13b and CasRx,respectively.Further analysis demonstrated that both PspCas13b and CasRx effectively down-regulated the transcription of exogenouslyintroduced target and endogenous genes in these cell lines.In addition,we generated two transgenic silkworm strains,one expressing CasRx and the other expressing RNA-guided CRISPR RNA targeting Sex combs reduced(Scr).Further crossing experiments showed that CasRx induced a down-regulation of Scr transcription in silkworms,which impaired systemic growth of larvae.Overall,this study demonstrated that the CRISPR-Cas13RNA editing system works efficiently in the silkworm,providing a potential alternative approach for RNA manipulation in lepidopteran insects.展开更多
DEAR EDITOR,Based upon morphological and molecular evidence,the authors revised the genus Rohanixalus Biju,Garg,Gokulakrishnan,Chandrakasan,Thammachoti,Ren,Gopika,Bisht,Hamidy and Shouche,2020(Anura:Rhacophoridae)in C...DEAR EDITOR,Based upon morphological and molecular evidence,the authors revised the genus Rohanixalus Biju,Garg,Gokulakrishnan,Chandrakasan,Thammachoti,Ren,Gopika,Bisht,Hamidy and Shouche,2020(Anura:Rhacophoridae)in China through describing one new species,adding one species to the fauna(R.shyamrupus)and supplementing data on one species(Rohanixalus hansenae;Supplementary Materials).展开更多
AlphaFold3(AF3),as the latest generation of artificial intelligence model jointly developed by Google DeepMind and Isomorphic Labs,has been widely heralded in the scientific research community since its launch.With un...AlphaFold3(AF3),as the latest generation of artificial intelligence model jointly developed by Google DeepMind and Isomorphic Labs,has been widely heralded in the scientific research community since its launch.With unprecedented accuracy,the AF3 model may successfully predict the structure and interactions of virtually all biomolecules,including proteins,ligands,nucleic acids,ions,etc.By accurately simulating the structural information and interactions of biomacromolecules,it has shown great potential in many aspects of structural prediction,mechanism research,drug design,protein engineering,vaccine development,and precision therapy.In order to further understand the characteristics of AF3 and accelerate its promotion,this article sets out to address the development process,working principle,and application in drugs and biomedicine,especially focusing on the intricate differences and some potential pitfalls compared to other deep learning models.We explain how a structure-prediction tool can impact many research fields,and in particular revolutionize the strategies for designing of effective next generation vaccines and chemical and biological drugs.展开更多
Respiration is a vital process essential for organism survival,with most terrestrial insects relying on a sophisticated tubular tracheal network.In the current study,a gene with repetitive sequence was identified with...Respiration is a vital process essential for organism survival,with most terrestrial insects relying on a sophisticated tubular tracheal network.In the current study,a gene with repetitive sequence was identified within the silkworm genome.Designated as BmMuc91C,it contains a dozen repeated motifs“PSSSYGAPX”and“GGYSSGGX”in its sequence.BmMuc91C exhibits specific expression in the tracheal system of silkworm larvae,with significantly higher expression levels during the molting stage.Overexpression of BmMuc91C in individual silkworms resulted in a marked increase in tracheal diameter,particularly during the molting stage.Immunofluorescence staining using a BmMuc91C antibody revealed a noticeable thickening of the apical extracellular matrix in the trachea.Tensile testing confirmed a considerable enhancement in tracheal elasticity.Additionally,a BmMuc91C mutation strain of silkworms was generated using the clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated nuclease 9 system.Although no significant differences were observed in the growth,development,and molting of BmMuc91C mutant silkworms,mechanical tests demonstrated a decrease in tracheal elasticity.Transcriptomic techniques revealed that a significant number of cuticular and chitin-binding proteins were among the differentially expressed genes between mutant and wild-type silkworms.Furthermore,the recombined BmMuc91C protein was successfully expressed using the Escherichia coli system.Cross-linking experiments with horseradish peroxidase demonstrated the formation of macromolecular complexes of BmMuc91C,which exhibited spontaneous luminescent properties under ultraviolet light.This research sheds light on the role of elastic proteins in insect tracheae and provides valuable insights for the development of elastic biomaterials.展开更多
Alternating hemiplegia of childhood(AHC)is an infrequent genetic pathology manifesting primarily as a neurodevelopmental disorder[1].Affected infants exhibit recurrent paroxysmal manifestations within several months p...Alternating hemiplegia of childhood(AHC)is an infrequent genetic pathology manifesting primarily as a neurodevelopmental disorder[1].Affected infants exhibit recurrent paroxysmal manifestations within several months postpartum,including hemiplegia,dystonia,abnormal ocular movements,and seizures[2].Additionally,patients may present with persistent hypotonia,developmental delay,and cognitive impairments[2].Approximately 70%of AHC cases are linked to deleterious mutations in the ATP1A3 gene[3],which encodes theα3 subunit of the sodium–potassium pump(Na+/K+ATPase),essential for neuronal homeostasis[4].Dysfunction of ATP1A3 activity can lead to neuronal hyperexcitability or energy metabolism imbalances,resulting in episodic neurological symptoms and developmental anomalies[5].Furthermore,ATP1A3 mutations with D801N,E815K,and G947R account for>65%of AHC cases,with incidences of 36%,22%,and 9%,respectively[1].Previous studies have revealed a dominant-negative pathogenic mechanism associated with AHC-related ATP1A3 mutations,wherein the aberrant ATP1A3 protein not only loses its intrinsic functionality but also impedes the functionality of other proteins[4].This distinct pathological paradigm has posed significant challenges to the advancement of conventional gene therapy approaches.Currently,there are no curative or highly effective treatments available for this disease.展开更多
Chronic non-healing wounds,such as diabetic foot,pressure sores and bedsores have seriously affected the life quality of patients worldwide.GFs provide a potential solution to promote chronic wound healing by promotin...Chronic non-healing wounds,such as diabetic foot,pressure sores and bedsores have seriously affected the life quality of patients worldwide.GFs provide a potential solution to promote chronic wound healing by promoting cell proliferation and differentiation.However,limited resource,high cost,and instability in vivo greatly hin-dered their clinical applications.In present study,two silk gland bioreactor silkworm stains were generated to successfully synthesize functional silk fibers incorporating high expressions of EGF and PDGF-BB.The two GFs functionalized silk raw materials were used to fabricate a dual GFs sericin hydrogel(E/P-SH)delivering system with tunable material performances for better cell adhesion and biocompatibility,sustainable release of the dual GFs to synergistically promote cell proliferation and migration,which realized the significant healing of chronic full-thickness skin wound in diabetic mice within 12 days with more organized collagen arrangement and better epithelialization degree by reducing inflammatory response and promoting vascularization.These findings demonstrated that the biosynthesized dual GFs-SH delivering system provides an opportunity to broaden the wide utility of GFs in clinical treatment of diabetic wound healing.展开更多
Ciliary neurotrophic factor(CNTF)acts as a potent neuroprotective agent in neuronal survival and regeneration,and can also induce the differentiation of several stem cells into neurons,which highlights the broad appli...Ciliary neurotrophic factor(CNTF)acts as a potent neuroprotective agent in neuronal survival and regeneration,and can also induce the differentiation of several stem cells into neurons,which highlights the broad application of CNTF in biomedicine.However,large-scale production of bioactive recombinant human CNTF protein remains to be explored.Herein,this study aims to express a bioactive human CNTF protein on a large scale by genetically engineering a silk gland bioreactor of silkworm.Our results showed that CNTF protein was successfully expressed in the middle silk gland(MSG)of silkworm,which can be secreted into the silks with the amount of 3.2 mg/g cocoons.The fabrication of human CNTF-functionalized silk material was able to promote proliferation and migration of neural cells when compared to the natural silk protein.Importantly,this functional silk material could also facilitate neurite outgrowth of mouse retinal ganglion cell(RGC-5)cells.All these data demonstrated a high bioactivity of the recombinant human CNTF protein expressed in the MSG of silkworm.The further fabrication of different silk materials with CNTF bioactivity will give biomedical applications in tissue engineering and neuroregeneration.展开更多
An increasing body of evidence shows that the lipid droplet,a neutral lipid storage organelle,plays a role in lipid metabolism and energy homeostasis through its interaction with mitochondria.However,the cellular func...An increasing body of evidence shows that the lipid droplet,a neutral lipid storage organelle,plays a role in lipid metabolism and energy homeostasis through its interaction with mitochondria.However,the cellular functions and molecular mechanisms of the interaction remain ambiguous.Here we present data from transmission electron microscopy,fluorescence imaging,and reconstitution assays,demonstrating that lipid droplets physically contact mitochondria in vivo and in vitro.Using a bimolecular fluorescence complementation assay in Saccharomyces cerevisiae,we generated an interactomic map of protein-protein contacts of lipid droplets with mitochondria and peroxisomes.The lipid droplet proteins Erg6 and Pet10 were found to be involved in 75%of the interactions detected.Interestingly,interactions between 3 pairs of lipid metabolic enzymes were detected.Collectively,these data demonstrate that lipid droplets make physical contacts with mitochondria and peroxisomes,and reveal specific molecular interactions that suggest active participation of lipid droplets in lipid metabolism in yeast.展开更多
Prime editing(PE)is a versatile CRISPR-Cas based precise genome-editing platform widely used to introduce a range of possible base conversions in various organisms.However,no PE systems have been shown to induce herit...Prime editing(PE)is a versatile CRISPR-Cas based precise genome-editing platform widely used to introduce a range of possible base conversions in various organisms.However,no PE systems have been shown to induce heritable mutations in tobacco,nor in any other dicot.In this study,we generated an efficient PE system in tobacco that not only introduced heritable mutations,but also enabled anthocyanin-based reporter selection of transgene-free T_(1) plants.This system was used to confer Zabienol biosynthesis in the allotetraploid tobacco cultivar HHDJY by restoring a G>T conversion in the NtCPS2 gene.High levels of Z-abienol were detected in the leaves of homozygous T_(1) plants at two weeks after topping.This study describes an advance in PE systems and expands genome-editing toolbox in tobacco,even in dicots,for use in basic research and molecular breeding.And restoring biosynthesis of Z-abienol in tobacco might provide an efficient way to obtain Z-abienol in plants.展开更多
Mycophenolic acid(MPA,1)and its derivatives are first-line immunosuppressants used in organ transplantation and for treating autoimmune diseases.Despite chemical synthetic achievements,the biosynthetic formation of a ...Mycophenolic acid(MPA,1)and its derivatives are first-line immunosuppressants used in organ transplantation and for treating autoimmune diseases.Despite chemical synthetic achievements,the biosynthetic formation of a seven-carbon carboxylic acid pharmacophore side chain of 1,especially the processes involving the cleavage of the prenyl side chain between DHMP(4)and DMMPA(5),remains unknown.In this work,we identified a membrane-bound prenyltransferase,PgMpaA,that transfers FPP to 4 to yield FDHMP(6).Compound 6 undergoes the first cleavage step via a new globin-like enzyme PgMpaB to form a cryptic intermediate 12.Heterologous expression of PgMpa genes in Aspergillus nidulans demonstrates that the second cleavage step(from 12 to 5)of 1 is a PgMpa clusterindependent process in vivo.Our results,especially the discovery of the broad tolerance of substrates recognized by PgMpaB,set up a strategy for the formation of"pseudo-isopentenyl"natural products using fungal globin-like enzymes.展开更多
Extracellular superoxide dismutase(EcSOD)protects tissues from oxidative stress,and thus is considered as a therapeutic agent for many diseases such as atherosclerosis,hypertension,and cancer.However,cost-effective pr...Extracellular superoxide dismutase(EcSOD)protects tissues from oxidative stress,and thus is considered as a therapeutic agent for many diseases such as atherosclerosis,hypertension,and cancer.However,cost-effective production of bioactive recombinant human EcSOD(rhEcSOD)remains a challenge.Herein,we developed an efficient strategy for producing active rhEcSOD by transgenic silkworms.rhEcSOD was successfully synthesized as homodimers and homotetramers in the middle silk gland and spun into the cocoons with a concentration of 9.48±0.21 mg/g.Purification of rhEcSOD from the cocoons could be conveniently achieved with a purity of 99.50%and a yield of 3.5±0.5 mg/g.Additionally,N-glycosylation at the only site of N89 in rhEcSOD with 10 types were identified.The purified rhEcSOD gained the potent enzymatic activity of 4162±293 U/mg after Cu/Zn ions incorporation.More importantly,rhEcSOD was capable of penetrating and accumulating in the nuclei of cells to maintain cell morphology and attenuate ultraviolet B-induced cell apoptosis by eliminating reactive oxygen species and inhibiting the C-Jun N-terminal kinase signaling pathway.These results demonstrated that the transgenic silkworm could successfully produce rhEcSOD with enzymatic and biological activities for biomedical applications.展开更多
MicroRNAs play critical roles in multiple developmental processes in insects.Our previous study showed that CRISPR/Cas9-mediated knock down of the microRNA let-7 in silkworms increased the size of larvae and silk glan...MicroRNAs play critical roles in multiple developmental processes in insects.Our previous study showed that CRISPR/Cas9-mediated knock down of the microRNA let-7 in silkworms increased the size of larvae and silk glands,thereby improving the silk production capacity.In this study,we elucidate the molecular mechanism underlying of let-7 regulates growth.Identification of differentially expressed genes in response to let-7 knock down revealed enrichment of pathways associated with cell proliferation and DNA replication.let-7 dysregulation affected the cell cycle and proliferation of the Bombyx mori cell line BmN.Dual-luciferase and target site mutation assays showed that BmCDK1 is a direct target gene of let-7,with only 1 binding site on its 3′-untranslated region.RNA interference of BmCDK1 inhibited cell proliferation,but this effect was counteracted by co-transfection with let-7 antagomir.Moreover,let-7 knock down induced BmCDK1 expression and promoted cell proliferation in multiple tissues,and further induced endomitosis in the silk gland in vivo.Knock down of BmCDK1 resulted in abnormal formation of a new epidermis,and larval development was arrested at the 2nd or 3rd molt stage.Taken together,our results demonstrated that BmCDK1 is a novel target of let-7 in cell fate determination,possessing potential for improving silk yield in silkworm.展开更多
Clip-domain serine proteases (CLIPs) play important roles in insect innate immunity and development. Our previous studies indicated that CLIP13, an epidermis-specific gene, was involved in cuticle remodeling during mo...Clip-domain serine proteases (CLIPs) play important roles in insect innate immunity and development. Our previous studies indicated that CLIP13, an epidermis-specific gene, was involved in cuticle remodeling during molting and metamorphosis in the silkworm, Bombyx mori. However, the transcriptional regulatory mechanism and regulatory pathways of CLIP13 remained unclear. In the present study, we investigated CLIP13 expression and the regulation pathway controlled by 20-hydroxyecdysone (20E) in the silkworm. At the transcriptional level, expression of CLIP13 exhibited pronounced spatial and temporal specificity in different regions of the epidermis;homeodomain transcription factors POU-M2, antennapedia (Antp), and abdominal-B (Abd-B) showed similar expression change trends as CLIP13 in the head capsule, thorax, and abdomen, respectively. Furthermore, results of cell transfection assays, electrophoretic mobility shift assays, and chromatin immunoprecipitation demonstrated that POU-M2, Antp, and Abd-B were involved in the transcriptional regulation of CLIP13 by directly binding to their cis-response elements in CLIP13 promoter. RNA interference-mediated silencing of POU-M2, Antp, and Abd-B led to a decrease of CLIP13 expression in the head capsule, the epidermis of the 1st to 3rd thoracic segments and the 7th to 10th abdominal segments, respectively. Consistent with CLIP13, 20E treatment significantly upregulated expression of POU-M2, Antp, and Abd-B in the silkworm epidermis. Taken together, these data suggest that 20E positively regulates transcription of CLIP13 via homeodomain proteins POU-M2, Antp, and Abd-B in different regions of the silkworm epidermis during metamorphosis, thus affecting the molting process. Our findings provide new insight into the functions of homeodomain transcription factors in insect molting.展开更多
Clustered regularly interspaced short palindromic repeats(CRISPR)-associated systems(Cas)are efficient tools for targeting specific genes for laboratory research,agricultural engineering,biotechnology,and human diseas...Clustered regularly interspaced short palindromic repeats(CRISPR)-associated systems(Cas)are efficient tools for targeting specific genes for laboratory research,agricultural engineering,biotechnology,and human disease treatment.Cas9,by far the most extensively used gene-editing nuclease,has shown great promise for the treatment of hereditary diseases,viral infection,cancers,and so on.Recent reports have revealed that some other types of CRISPR-Cas systems may also have surprising potential to join the fray as gene-editing tools for various applications.Despite the rapid progress in basic research and clinical tests,some underlying problems present continuous,significant challenges,such as editing efficiency,relative difficulty in delivery,off-target effects,immunogenicity,etc.This article summarizes the applications of CRISPR-Cas from bench to bedside and highlights the current obstacles that may limit the usage of CRISPR-Cas systems as gene-editing toolkits in precision medicine and offer some viewpoints that may help to tackle these challenges and facilitate technical development.CRISPR-Cas systems,as a powerful gene-editing approach,will offer great hopes in clinical treatments for many individuals with currently incurable diseases.展开更多
Transcription factor Broad Complex(BR-C)is an ecdysone primary response gene in insects and participates in the regulation of insect growth and development.In this study,we performed a genome-wide identification of BR...Transcription factor Broad Complex(BR-C)is an ecdysone primary response gene in insects and participates in the regulation of insect growth and development.In this study,we performed a genome-wide identification of BR-C target genes in silkworm(Bombyx mori)using chromatin immunoprecipitation followed by high-throughput sequencing(ChIP-seq).As a result,a total of 1006 BR-C ChIP peaks were identified,and 15%of peaks were located in the promoter regions of 133 protein-coding genes.Functional annotation revealed that these ChIP peak-associated genes,as potential BR-C targets,were enriched in pathways related to biosynthetic process,metabolic process,and development.Transcriptome analysis and quantitative real-time polymerase chain reaction(PCR)examination revealed that developmental changes in expression patterns of a portion of potential BR-C targets,including HR96 and GC-otl,were similar to those of BR-C.ChIP-PCR examination confirmed that BR-C could directly bind to the promoters of potential targets.Further,dual luciferase assays demonstrated that HR96 promoter activity was significantly upregulated following BR-C overexpression,and this upregu-lation was abolished when the binding motif in the promoter was truncated.This study will be helpful for deciphering the regulatory roles of BR-C during insect growth and development.展开更多
基金supported by the Shenzhen Science and Technology Innovation Commission,No.JCYJ20220818103009018(to YD).
文摘The arginine-phenylalanine-amide neuropeptide receptor family comprises a subclass within the G protein-coupled receptor superfamily with crucial roles in physiological regulation.These receptors recognize and bind neuropeptides with an arginine-phenylalanine-amide motif,thereby participating in a variety of biological processes such as energy metabolism,pain perception,and reproductive functions.In this review,we explore the physiological and pathological processes involving these receptors and delve into the structure-activity relationships of their ligand peptides,clarifying the key structural motifs within these neuropeptides that determine their biological activity,pharmacological potency,and receptor selectivity.Particular emphasis is placed on their roles in modulating nociception,regulating appetite,and maintaining reproductive health.Additionally,we discuss the therapeutic potential of structure-based drug design targeting these receptors based on existing cryo-electron microscopy structures.The available structural insights into ligand-binding pockets and G protein-receptor interaction interfaces provide a clear perspective and valuable complement to ligand optimization.
基金funded by the National Natural Science Foundation of China(Nos.82201594,81502582)National Key Research Project of MOST(2023YFA0915000)+5 种基金Fund of the Affiliated Xiangshan Hospital of Wenzhou Medical University,and Wenzhou Institute University of Chinese Academy of SciencesFunding was also provided by the Fundamental Research Funds for the Central Universities(N182004002,N2220002)Natural Science Foundation of Liaoning Province(2021-MS-104,2022-YGJC-39,2022-MS-228)Fundamental Scientific Research Fund of Liaoning Provincial Education Department(LJKQZ2021002)supported by the Open project of State Key Laboratory of Animal Biotech Breeding(No.2025SKLAB6-13)Construction Project of Liaoning Provincial Key Laboratory,China(2022JH13/10200026).
文摘From a neuroscience perspective,cancer neuroscience has emerged as a subfield of cancer research.Presumable mechanisms underlying cancer-related neuronal activity(termed neurosciences)include the induction and modulation of signaling pathways that govern cell fate determination and emotional responses(anxiety and stress),such as structural molecules(synaptic structures and current transduction)and secretory substances(neurotransmitters,cytokines,hormones and neuropeptides).In the past 3 years,these neuronal activities,which can either promote cancer growth or be hijacked by cancer cells to support tumor survival and invasion,have been widely demonstrated to be closely related to cancer progression.The molecular mechanisms are also being refined.Despite their great promise,translating neuroscientific discoveries into clinically actionable strategies for cancer diagnosis,prognosis,and treatment remains a formidable task.In this comprehensive review,we attempt to provide a full account of the intersection between neuroscience and cancer research.From the perspective of cancer neuroscience,we fully discuss the potential signaling molecules and their regulatory mechanisms,as well as targets and emerging therapeutic strategies that control tumor progression via multiomics approaches.Overall,cancer neuroscience may have unprecedented potential for understanding neuronal functions and cancer development,ultimately offering the significantly improved cancer treatment.
基金financially supported by National Key R&D Program of China(No.2016YFD0400200)National Natural Science Foundation of China(No.31972102,31671881,and 31901683)+4 种基金Chongqing Research Program of Basic Research and Frontier Technology(No.cstc2018jcyj A0939)Chongqing Technology Innovation and Application Demonstration Project(No.cstc2018jscx-msyb X0204)Fundamental Research Funds for the Central Universities(No.XDJK2019B028)Innovation Program for Chongqing’s Overseas Returnees(cx2019072)Fundamental Research Funds for the Central Universities,China(SWU 019009)。
文摘Ultraviolet(UV)-induced photoaging skin has become an urgent issue.The functional foods and cosmetics aiming to improve skin photoaging are developing rapidly,and the demand is gradually increasing year by year.Collagen peptides have been proven to display diverse physiological activities,such as excellent moisture retention activity,hygroscopicity,tyrosinase inhibitory activity and antioxidant activity,which indicates that they have great potential in amelioration of UV-induced photoaging.The main objective of this article is to recap the main mechanisms to improve photoaging skin by collagen peptides and their physiological activities in photo-protection.Furthermore,the extraction and structural characteristics of collagen peptides are overviewed.More importantly,some clinical trials on the beneficial effect on skin of collagen peptides are also discussed.In addition,prospects and challenges of collagen peptides are emphatically elucidated in this review.This article implies that collagen peptides have great potential as an effective ingredient in food and cosmetics industry with a wide application prospect.
基金supported by a Grant-in-Aid for Young Scientists B 23792391 from the Ministry of Education,Culture,Sports, Science and Technology,Japan
文摘In the present study,we investigate the expression profile of the epidermal growth factor receptor family,which comprises EGFR/ ErbBl,HER2/ErbB2,HER3/ErbB3 and HER4/ErbB4 in oral leukoplakia(LP).The expression of four epidermal growth factor receptor (EGFR) family genes and their ligands were measured in LP tissues from 14 patients and compared with levels in 10 patients with oral lichen planus(OLP) and normal oral mucosa(NOM) from 14 healthy donors by real-time polymerase chain reaction(PCR) and immunohistochemistry.Synchronous mRNA coexpression of ErbBl,ErbB2,ErbB3and ErbB4 was detected in LP lesions.Out of the receptors,only ErbB4 mRNA and protein was more highly expressed in LP compared with NOM tissues.These were strongly expressed by epithelial keratinocytes in LP lesions,as shown by immunohistochemistry.Regarding the ligands,the mRNA of Neuregulin2 and 4 were more highly expressed in OLP compared with NOM tissues.Therefore,enhanced ErbB4 on the keratinocytes and synchronous modulation of EGFR family genes may contribute to the pathogenesis and carcinogenesis of LP.
基金This work was supported by funding from the Ginger Genome Project of Chongqing University of Arts and Sciences(2018)the Natural Science Foundation of Chongqing(cstc2019jcyj-msxmX0300,cstc2019jcyj-msxmX0697,CQYC201903201,cstc2019jscx-dxwtBX0028)+4 种基金the Foundation for High-level Talents of Chongqing University of Arts and Science(2017RTZ21,P2018TZ05)the Scientific and Technological Research Program of Chongqing Municipal Education Commission(KJZD-K202001304,KJQN201801339,KJQN201801330,KJQN201801335)the Foundation of Hubei Rural Science and Technology(2020BBA037)the State Key Research and Development Program of Hubei(2020BBA037)the Foundation of Laiwu Experimental Station of the National Characteristic Vegetable Industry System.
文摘Ginger(Zingiber officinale),the type species of Zingiberaceae,is one of the most widespread medicinal plants and spices.Here,we report a high-quality,chromosome-scale reference genome of ginger‘Zhugen’,a traditionally cultivated ginger in Southwest China used as a fresh vegetable,assembled from PacBio long reads,Illumina short reads,and high-throughput chromosome conformation capture(Hi-C)reads.The ginger genome was phased into two haplotypes,haplotype 1(1.53 Gb with a contig N50 of 4.68 M)and haplotype 0(1.51 Gb with a contig N50 of 5.28 M).Homologous ginger chromosomes maintained excellent gene pair collinearity.In 17,226 pairs of allelic genes,11.9%exhibited differential expression between alleles.Based on the results of ginger genome sequencing,transcriptome analysis,and metabolomic analysis,we proposed a backbone biosynthetic pathway of gingerol analogs,which consists of 12 enzymatic gene families,PAL,C4H,4CL,CST,C3’H,C3OMT,CCOMT,CSE,PKS,AOR,DHN,and DHT.These analyses also identified the likely transcription factor networks that regulate the synthesis of gingerol analogs.Overall,this study serves as an excellent resource for further research on ginger biology and breeding,lays a foundation for a better understanding of ginger evolution,and presents an intact biosynthetic pathway for species-specific gingerol biosynthesis.
基金supported by the National Natural Science Foundation of China(32070496,32370555)Fundamental Research Funds for the Central Universities(SWU120033)Technology Innovation and Application Development Program of Chongqing(CSTB2024TIADKPX0023)。
文摘The CRISPR-Cas13 system,an RNA-guided editing tool,has emerged as a highly efficient and stable RNA editing technique.Although the CRISPR-Cas13 system has been developed in several insect species,its application in lepidopterans has not yet been reported.In the present study,we evaluated the RNA cleavage activity of the CRISPR-Cas13 system in the silkworm(Bombyx mori),a model lepidopteran insect,both ex vivo and in vivo.We established two stable silkworm BmE cell lines expressing PspCas13b and CasRx,respectively.Further analysis demonstrated that both PspCas13b and CasRx effectively down-regulated the transcription of exogenouslyintroduced target and endogenous genes in these cell lines.In addition,we generated two transgenic silkworm strains,one expressing CasRx and the other expressing RNA-guided CRISPR RNA targeting Sex combs reduced(Scr).Further crossing experiments showed that CasRx induced a down-regulation of Scr transcription in silkworms,which impaired systemic growth of larvae.Overall,this study demonstrated that the CRISPR-Cas13RNA editing system works efficiently in the silkworm,providing a potential alternative approach for RNA manipulation in lepidopteran insects.
基金supported by the Fundamental Research Fund for Central Universities (SWU-KR22014)National Natural Science Foundation of China (NSFC32170478,32370478)+8 种基金Yunnan Fundamental Research Project (202001AW070016,202005AC160046)“Special Fund for Youth Team of Southwest University” (SWU-XJPY202302)to Y.Z.Y.National Key R&D Program of China (2022YFC2602500)the Second Tibetan Plateau Scientific Expedition and Research Program (STEP) (2019QZKK0501)Survey of Wildlife Resources in Key Areas of Xizang (ZL202203601)China’s Biodiversity Observation Network (Sino-BON)Animal Branch of Germplasm Bank of Wild Species,Chinese Academy of Sciences (Large Research Infrastructure Fund)to J.C.Unit of Excellence 2024 on Integrative diversity assessment of aquatic animals from Thailand (Fundamental FundFF67)to C.S。
文摘DEAR EDITOR,Based upon morphological and molecular evidence,the authors revised the genus Rohanixalus Biju,Garg,Gokulakrishnan,Chandrakasan,Thammachoti,Ren,Gopika,Bisht,Hamidy and Shouche,2020(Anura:Rhacophoridae)in China through describing one new species,adding one species to the fauna(R.shyamrupus)and supplementing data on one species(Rohanixalus hansenae;Supplementary Materials).
基金supported by funds of the Key Research and Development Grant of MOST(grant No.2023YFA095000)the Affiliated Xiangshan Hospital of Wenzhou Medical University,the National Science Foundation of China(grant No.82470005)Wenzhou Institute University of Chinese Academy of Sciences.
文摘AlphaFold3(AF3),as the latest generation of artificial intelligence model jointly developed by Google DeepMind and Isomorphic Labs,has been widely heralded in the scientific research community since its launch.With unprecedented accuracy,the AF3 model may successfully predict the structure and interactions of virtually all biomolecules,including proteins,ligands,nucleic acids,ions,etc.By accurately simulating the structural information and interactions of biomacromolecules,it has shown great potential in many aspects of structural prediction,mechanism research,drug design,protein engineering,vaccine development,and precision therapy.In order to further understand the characteristics of AF3 and accelerate its promotion,this article sets out to address the development process,working principle,and application in drugs and biomedicine,especially focusing on the intricate differences and some potential pitfalls compared to other deep learning models.We explain how a structure-prediction tool can impact many research fields,and in particular revolutionize the strategies for designing of effective next generation vaccines and chemical and biological drugs.
基金supportedbytheNational Key Research and Development ProgramofChina(2022YFD1201600)the Program of NationaNl atural Science of China(No.32030103)Technology Innovation andApplication Development Program of Chongqing(CSTB2024TIAD-KPX0023,CSTB2024TIAD-KPX0026)。
文摘Respiration is a vital process essential for organism survival,with most terrestrial insects relying on a sophisticated tubular tracheal network.In the current study,a gene with repetitive sequence was identified within the silkworm genome.Designated as BmMuc91C,it contains a dozen repeated motifs“PSSSYGAPX”and“GGYSSGGX”in its sequence.BmMuc91C exhibits specific expression in the tracheal system of silkworm larvae,with significantly higher expression levels during the molting stage.Overexpression of BmMuc91C in individual silkworms resulted in a marked increase in tracheal diameter,particularly during the molting stage.Immunofluorescence staining using a BmMuc91C antibody revealed a noticeable thickening of the apical extracellular matrix in the trachea.Tensile testing confirmed a considerable enhancement in tracheal elasticity.Additionally,a BmMuc91C mutation strain of silkworms was generated using the clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated nuclease 9 system.Although no significant differences were observed in the growth,development,and molting of BmMuc91C mutant silkworms,mechanical tests demonstrated a decrease in tracheal elasticity.Transcriptomic techniques revealed that a significant number of cuticular and chitin-binding proteins were among the differentially expressed genes between mutant and wild-type silkworms.Furthermore,the recombined BmMuc91C protein was successfully expressed using the Escherichia coli system.Cross-linking experiments with horseradish peroxidase demonstrated the formation of macromolecular complexes of BmMuc91C,which exhibited spontaneous luminescent properties under ultraviolet light.This research sheds light on the role of elastic proteins in insect tracheae and provides valuable insights for the development of elastic biomaterials.
基金funded by the National Natural Science Foundation of China(grant Nos.32470022 and 82020108021)Chongqing Outstanding Youth Funds(grant No.CSTB2025NSCQ-JQX)+1 种基金the National Natural Science Foundation of Chongqing(grant No.cstc2024ycjh-bgzxm0076)the Fundamental Research Funds for the Central Universities(grant No.SWU-KR22013).
文摘Alternating hemiplegia of childhood(AHC)is an infrequent genetic pathology manifesting primarily as a neurodevelopmental disorder[1].Affected infants exhibit recurrent paroxysmal manifestations within several months postpartum,including hemiplegia,dystonia,abnormal ocular movements,and seizures[2].Additionally,patients may present with persistent hypotonia,developmental delay,and cognitive impairments[2].Approximately 70%of AHC cases are linked to deleterious mutations in the ATP1A3 gene[3],which encodes theα3 subunit of the sodium–potassium pump(Na+/K+ATPase),essential for neuronal homeostasis[4].Dysfunction of ATP1A3 activity can lead to neuronal hyperexcitability or energy metabolism imbalances,resulting in episodic neurological symptoms and developmental anomalies[5].Furthermore,ATP1A3 mutations with D801N,E815K,and G947R account for>65%of AHC cases,with incidences of 36%,22%,and 9%,respectively[1].Previous studies have revealed a dominant-negative pathogenic mechanism associated with AHC-related ATP1A3 mutations,wherein the aberrant ATP1A3 protein not only loses its intrinsic functionality but also impedes the functionality of other proteins[4].This distinct pathological paradigm has posed significant challenges to the advancement of conventional gene therapy approaches.Currently,there are no curative or highly effective treatments available for this disease.
基金supported by the National Key Research and Devel-opment Program of China(2022YFD1201600)Chongqing Science and Technology Commission(CSTB2024NSCQ-QCXMX0017)National Natural Science Foundation of China(32030103).
文摘Chronic non-healing wounds,such as diabetic foot,pressure sores and bedsores have seriously affected the life quality of patients worldwide.GFs provide a potential solution to promote chronic wound healing by promoting cell proliferation and differentiation.However,limited resource,high cost,and instability in vivo greatly hin-dered their clinical applications.In present study,two silk gland bioreactor silkworm stains were generated to successfully synthesize functional silk fibers incorporating high expressions of EGF and PDGF-BB.The two GFs functionalized silk raw materials were used to fabricate a dual GFs sericin hydrogel(E/P-SH)delivering system with tunable material performances for better cell adhesion and biocompatibility,sustainable release of the dual GFs to synergistically promote cell proliferation and migration,which realized the significant healing of chronic full-thickness skin wound in diabetic mice within 12 days with more organized collagen arrangement and better epithelialization degree by reducing inflammatory response and promoting vascularization.These findings demonstrated that the biosynthesized dual GFs-SH delivering system provides an opportunity to broaden the wide utility of GFs in clinical treatment of diabetic wound healing.
基金supported by National Key Research and Development Program of China(No.2022YFD1201600)National Natural Science Foundation of China(Nos.32030103 and 32172798)+1 种基金Natural Science Foundation of Chongqing(Nos.cstc2020jcyjcxttX0001andCSTB2023NSCQ-MSX0814)Innovation and Entrepreneurship Training Program of Southwest University(No.S202210635112)。
文摘Ciliary neurotrophic factor(CNTF)acts as a potent neuroprotective agent in neuronal survival and regeneration,and can also induce the differentiation of several stem cells into neurons,which highlights the broad application of CNTF in biomedicine.However,large-scale production of bioactive recombinant human CNTF protein remains to be explored.Herein,this study aims to express a bioactive human CNTF protein on a large scale by genetically engineering a silk gland bioreactor of silkworm.Our results showed that CNTF protein was successfully expressed in the middle silk gland(MSG)of silkworm,which can be secreted into the silks with the amount of 3.2 mg/g cocoons.The fabrication of human CNTF-functionalized silk material was able to promote proliferation and migration of neural cells when compared to the natural silk protein.Importantly,this functional silk material could also facilitate neurite outgrowth of mouse retinal ganglion cell(RGC-5)cells.All these data demonstrated a high bioactivity of the recombinant human CNTF protein expressed in the MSG of silkworm.The further fabrication of different silk materials with CNTF bioactivity will give biomedical applications in tissue engineering and neuroregeneration.
基金supported by grants from the National Basic Research Program of China(Grant Nos.2009CB919000 and 2010CB833703)the National Natural Science Foundation of China(Grant Nos.30871229 and 30971431)the 21C Frontier Functional Proteomics Project(FPR08A1-060)funded by the Ministry of Education,Science and Technology,Republic of Korea.
文摘An increasing body of evidence shows that the lipid droplet,a neutral lipid storage organelle,plays a role in lipid metabolism and energy homeostasis through its interaction with mitochondria.However,the cellular functions and molecular mechanisms of the interaction remain ambiguous.Here we present data from transmission electron microscopy,fluorescence imaging,and reconstitution assays,demonstrating that lipid droplets physically contact mitochondria in vivo and in vitro.Using a bimolecular fluorescence complementation assay in Saccharomyces cerevisiae,we generated an interactomic map of protein-protein contacts of lipid droplets with mitochondria and peroxisomes.The lipid droplet proteins Erg6 and Pet10 were found to be involved in 75%of the interactions detected.Interestingly,interactions between 3 pairs of lipid metabolic enzymes were detected.Collectively,these data demonstrate that lipid droplets make physical contacts with mitochondria and peroxisomes,and reveal specific molecular interactions that suggest active participation of lipid droplets in lipid metabolism in yeast.
基金supported by Beijing Scholars Program (BSP041)Financial Special Fund of Beijing Academy of Agriculture and Forestry Sciences (CZZJ202206)+1 种基金the key projects of YNZY (2022JY02)CNTC (110202101034,JY-11)。
文摘Prime editing(PE)is a versatile CRISPR-Cas based precise genome-editing platform widely used to introduce a range of possible base conversions in various organisms.However,no PE systems have been shown to induce heritable mutations in tobacco,nor in any other dicot.In this study,we generated an efficient PE system in tobacco that not only introduced heritable mutations,but also enabled anthocyanin-based reporter selection of transgene-free T_(1) plants.This system was used to confer Zabienol biosynthesis in the allotetraploid tobacco cultivar HHDJY by restoring a G>T conversion in the NtCPS2 gene.High levels of Z-abienol were detected in the leaves of homozygous T_(1) plants at two weeks after topping.This study describes an advance in PE systems and expands genome-editing toolbox in tobacco,even in dicots,for use in basic research and molecular breeding.And restoring biosynthesis of Z-abienol in tobacco might provide an efficient way to obtain Z-abienol in plants.
基金supported by the National Natural Science Foundation of China(31870022)the Fundamental Research Funds for the Central Universities(XDJK2018B035 and 201941001,China)+3 种基金the Scientific Research Starting Foundation of Southwest University(SWU117034,China)the Marine S&T Fund of Shandong Province for Pilot National Laboratory for Marine Science and Technology(Qingdao,China)(No.2018SDKJ0401-2)Taishan Scholar Youth Expert Program in Shandong Province(tsqn201812021,China)supported by the Venture&Innovation Support Program for Chongqing Overseas Returnees and the Thousand Young Talents Program of China.
文摘Mycophenolic acid(MPA,1)and its derivatives are first-line immunosuppressants used in organ transplantation and for treating autoimmune diseases.Despite chemical synthetic achievements,the biosynthetic formation of a seven-carbon carboxylic acid pharmacophore side chain of 1,especially the processes involving the cleavage of the prenyl side chain between DHMP(4)and DMMPA(5),remains unknown.In this work,we identified a membrane-bound prenyltransferase,PgMpaA,that transfers FPP to 4 to yield FDHMP(6).Compound 6 undergoes the first cleavage step via a new globin-like enzyme PgMpaB to form a cryptic intermediate 12.Heterologous expression of PgMpa genes in Aspergillus nidulans demonstrates that the second cleavage step(from 12 to 5)of 1 is a PgMpa clusterindependent process in vivo.Our results,especially the discovery of the broad tolerance of substrates recognized by PgMpaB,set up a strategy for the formation of"pseudo-isopentenyl"natural products using fungal globin-like enzymes.
基金supported by the National Key Research and Development Program of China(2022YFD1201600)Chongqing Science and Technology Commission(cstc2020jcyj-cxttX0001,CSTB2022TIAD-KPX0083)Yongchuan District Science and Technology Commission(2022ycyfjg20002).
文摘Extracellular superoxide dismutase(EcSOD)protects tissues from oxidative stress,and thus is considered as a therapeutic agent for many diseases such as atherosclerosis,hypertension,and cancer.However,cost-effective production of bioactive recombinant human EcSOD(rhEcSOD)remains a challenge.Herein,we developed an efficient strategy for producing active rhEcSOD by transgenic silkworms.rhEcSOD was successfully synthesized as homodimers and homotetramers in the middle silk gland and spun into the cocoons with a concentration of 9.48±0.21 mg/g.Purification of rhEcSOD from the cocoons could be conveniently achieved with a purity of 99.50%and a yield of 3.5±0.5 mg/g.Additionally,N-glycosylation at the only site of N89 in rhEcSOD with 10 types were identified.The purified rhEcSOD gained the potent enzymatic activity of 4162±293 U/mg after Cu/Zn ions incorporation.More importantly,rhEcSOD was capable of penetrating and accumulating in the nuclei of cells to maintain cell morphology and attenuate ultraviolet B-induced cell apoptosis by eliminating reactive oxygen species and inhibiting the C-Jun N-terminal kinase signaling pathway.These results demonstrated that the transgenic silkworm could successfully produce rhEcSOD with enzymatic and biological activities for biomedical applications.
基金supported by grants from the National Natural Science Foundation of China(No.32102614)the National Key Research and Development Program of China(No.2022YFD1201600)+1 种基金the Natural Science Foundation of Chongqing,China(No.cstc2021jcyj-bsh0151)the Chongqing Special Support fund for Post Doctor(No.2010010006115189).
文摘MicroRNAs play critical roles in multiple developmental processes in insects.Our previous study showed that CRISPR/Cas9-mediated knock down of the microRNA let-7 in silkworms increased the size of larvae and silk glands,thereby improving the silk production capacity.In this study,we elucidate the molecular mechanism underlying of let-7 regulates growth.Identification of differentially expressed genes in response to let-7 knock down revealed enrichment of pathways associated with cell proliferation and DNA replication.let-7 dysregulation affected the cell cycle and proliferation of the Bombyx mori cell line BmN.Dual-luciferase and target site mutation assays showed that BmCDK1 is a direct target gene of let-7,with only 1 binding site on its 3′-untranslated region.RNA interference of BmCDK1 inhibited cell proliferation,but this effect was counteracted by co-transfection with let-7 antagomir.Moreover,let-7 knock down induced BmCDK1 expression and promoted cell proliferation in multiple tissues,and further induced endomitosis in the silk gland in vivo.Knock down of BmCDK1 resulted in abnormal formation of a new epidermis,and larval development was arrested at the 2nd or 3rd molt stage.Taken together,our results demonstrated that BmCDK1 is a novel target of let-7 in cell fate determination,possessing potential for improving silk yield in silkworm.
基金This work was supported by grants from the National Natural Science Foundation of China(31772532,31702187)the China Postdoctoral Science Foundation(2019M663880XB)Natural Science Foundation of Chongqing,China(cstc2019jcyj-bshX0033).
文摘Clip-domain serine proteases (CLIPs) play important roles in insect innate immunity and development. Our previous studies indicated that CLIP13, an epidermis-specific gene, was involved in cuticle remodeling during molting and metamorphosis in the silkworm, Bombyx mori. However, the transcriptional regulatory mechanism and regulatory pathways of CLIP13 remained unclear. In the present study, we investigated CLIP13 expression and the regulation pathway controlled by 20-hydroxyecdysone (20E) in the silkworm. At the transcriptional level, expression of CLIP13 exhibited pronounced spatial and temporal specificity in different regions of the epidermis;homeodomain transcription factors POU-M2, antennapedia (Antp), and abdominal-B (Abd-B) showed similar expression change trends as CLIP13 in the head capsule, thorax, and abdomen, respectively. Furthermore, results of cell transfection assays, electrophoretic mobility shift assays, and chromatin immunoprecipitation demonstrated that POU-M2, Antp, and Abd-B were involved in the transcriptional regulation of CLIP13 by directly binding to their cis-response elements in CLIP13 promoter. RNA interference-mediated silencing of POU-M2, Antp, and Abd-B led to a decrease of CLIP13 expression in the head capsule, the epidermis of the 1st to 3rd thoracic segments and the 7th to 10th abdominal segments, respectively. Consistent with CLIP13, 20E treatment significantly upregulated expression of POU-M2, Antp, and Abd-B in the silkworm epidermis. Taken together, these data suggest that 20E positively regulates transcription of CLIP13 via homeodomain proteins POU-M2, Antp, and Abd-B in different regions of the silkworm epidermis during metamorphosis, thus affecting the molting process. Our findings provide new insight into the functions of homeodomain transcription factors in insect molting.
基金This work was supported by the National Institutes of Health(NIH)(Grant No.AI138203-3)the American Association of Immunologists through a Careers in Immunology Fellowship.This work was also supported by the National Natural Science Foundation of China(Grants No.82020108021 and 32000033).
文摘Clustered regularly interspaced short palindromic repeats(CRISPR)-associated systems(Cas)are efficient tools for targeting specific genes for laboratory research,agricultural engineering,biotechnology,and human disease treatment.Cas9,by far the most extensively used gene-editing nuclease,has shown great promise for the treatment of hereditary diseases,viral infection,cancers,and so on.Recent reports have revealed that some other types of CRISPR-Cas systems may also have surprising potential to join the fray as gene-editing tools for various applications.Despite the rapid progress in basic research and clinical tests,some underlying problems present continuous,significant challenges,such as editing efficiency,relative difficulty in delivery,off-target effects,immunogenicity,etc.This article summarizes the applications of CRISPR-Cas from bench to bedside and highlights the current obstacles that may limit the usage of CRISPR-Cas systems as gene-editing toolkits in precision medicine and offer some viewpoints that may help to tackle these challenges and facilitate technical development.CRISPR-Cas systems,as a powerful gene-editing approach,will offer great hopes in clinical treatments for many individuals with currently incurable diseases.
基金funded by grants from the National Natural Science Foundation of China(31572464,31772679,and 32070496)the Natural Science Foundation of Chongqing(cstc2019jcyj-msxmX0446)the Fundamental Research Funds for the Central Universities(XDJK2020C008).
文摘Transcription factor Broad Complex(BR-C)is an ecdysone primary response gene in insects and participates in the regulation of insect growth and development.In this study,we performed a genome-wide identification of BR-C target genes in silkworm(Bombyx mori)using chromatin immunoprecipitation followed by high-throughput sequencing(ChIP-seq).As a result,a total of 1006 BR-C ChIP peaks were identified,and 15%of peaks were located in the promoter regions of 133 protein-coding genes.Functional annotation revealed that these ChIP peak-associated genes,as potential BR-C targets,were enriched in pathways related to biosynthetic process,metabolic process,and development.Transcriptome analysis and quantitative real-time polymerase chain reaction(PCR)examination revealed that developmental changes in expression patterns of a portion of potential BR-C targets,including HR96 and GC-otl,were similar to those of BR-C.ChIP-PCR examination confirmed that BR-C could directly bind to the promoters of potential targets.Further,dual luciferase assays demonstrated that HR96 promoter activity was significantly upregulated following BR-C overexpression,and this upregu-lation was abolished when the binding motif in the promoter was truncated.This study will be helpful for deciphering the regulatory roles of BR-C during insect growth and development.
