Soybean mosaic virus(SMV)poses a substantial threat to the yield and quality of soybean(Glycine max(L.)Merr.),leading to significant economic losses in soybean production.However,the mining of SMVresistance loci and t...Soybean mosaic virus(SMV)poses a substantial threat to the yield and quality of soybean(Glycine max(L.)Merr.),leading to significant economic losses in soybean production.However,the mining of SMVresistance loci and the exploration of the underlying disease resistance mechanisms remain relatively limited.MicroRNAs(miRNAs)are a class of post-transcriptional regulators that play a pivotal role in modulating plant growth,development and responding to various stresses.In this study,we demonstrated the function of the “miR398c/d-GmCSDs”module between soybean resistant and susceptible varieties,focusing on its differential regulatory roles in SMV infection.Specifically,SMV infection downregulated gma-miR398c/d expression in the resistant variety(Qihuang 1,QH),while upregulated them in the susceptible variety(Nannong 1138-2,NN).Transient expression assay in N.benthamiana confirmed that gma-miR398c/d can target six superoxide dismutase(SOD)family genes,which responded to SMV infection in both varieties.Stable overexpression of Gma-MIR398c/d in soybean or inhibition of the corresponding target genes’expression via Bean pod mottle virus(BPMV)-induced gene silencing(VIGS)led to reduced H_(2)O_(2)content and thereby promoted SMV infection.Conversely,plants overexpressing the target genes exhibited the opposite phenotypes.The functions of gma-miR398c/d and their target genes were further validated in N.benthamiana through transient co-expression with SMV infectious clone(pSC7-GFP),indicating that gma-miR398c/d negatively regulated soybean resistance to SMV,while the target genes positively contributed to disease resistance.Collectively,our findings provide novel insights into the regulatory mechanisms underlying soybean resistance to SMV.展开更多
Elevated lipoprotein(a)[Lp(a)]is a major independent risk factor for atheroscle-rotic cardiovascular disease(ASCVD),with limited response to traditional lipid-lowering therapies.Lepodisiran,a novel N-acetylgalactosami...Elevated lipoprotein(a)[Lp(a)]is a major independent risk factor for atheroscle-rotic cardiovascular disease(ASCVD),with limited response to traditional lipid-lowering therapies.Lepodisiran,a novel N-acetylgalactosamine-conjugated small interfering RNA,targets hepatic LPA message RNA to reduce apolipoprotein(a)production.Early-phase trials demonstrated>90%sustained Lp(a)reduction with excellent safety and tolerability.The phase 2 ALPACA trial confirmed dura-ble effects lasting up to one year after biannual dosing.Compared to other thera-pies,lepodisiran offers longer duration,high efficacy,and minimal side effects.Ongoing phase 3 studies aim to determine its impact on cardiovascular outcomes,potentially establishing a new standard in precise ASCVD risk management.展开更多
Objective: The mortality and morbidity rates associated with pancreatic cancer (PaCa) are extremely high. Various studies have demonstrated that pancreatic cancer will be the fourth cancer-related death by 2030, raisi...Objective: The mortality and morbidity rates associated with pancreatic cancer (PaCa) are extremely high. Various studies have demonstrated that pancreatic cancer will be the fourth cancer-related death by 2030, raising more concern for scholars to find effective methods to prevent and treat in order to improve the pancreatic cancer outcome. Using bioinformatic analysis, this study aims to pinpoint key genes that could impact PaCa patients’ prognosis and could be used as therapeutic targets. Methods: The TCGA and GEO datasets were integratively analyzed to identify prognosis-related differentially expressed genes. Next, the STRING database was used to develop PPI networks, and the MCODE and CytoNCA Cytoscape in Cytoscape were used to screen for critical genes. Through CytoNCA, three kinds of topology analysis were considered (degree, betweenness, and eigenvector). Essential genes were confirmed as potential target treatment through Go function and pathways enrichment analysis, a developed predictive risk model based on multivariate analysis, and the establishment of nomograms using the clinical information. Results: Overall, the GSE183795 and TCGA datasets associated 1311 and 2244 genes with pancreatic cancer prognosis, respectively. We identified 132 genes that were present in both datasets. The PPI network analysis using, the centrality analysis approach with the CytoNCA plug-in, showed that CDK2, PLK1, CCNB1, and TOP2A ranked in the top 5% across all three metrics. The independent analysis of a risk model revealed that the four key genes had a Hazard Ratio (HR) > 1. The monogram showed the predictive risk model and individual patient survival predictions were accurate. The results indicate that the effect of the selected vital genes was significant and that they could be used as biomarkers to predict a patient’s outcome and as possible target therapy in patients with pancreatic cancer. GO function and pathway analysis demonstrated that crucial genes might affect the P53 signaling pathway and FoxO signaling pathway, through which Meiotic nuclear division and cell cycle may have a significant function in essential genes affecting the outcome of patients who have pancreatic cancer. Conclusions: This study suggests that CDK2, CCNB1, PLK1 and TOP2A are four key genes that have a significant influence on PaCa migration and proliferation. CDK2, CCNB1, PLK1, and TOP2A can be used as potential PaCa prognostic biomarkers and therapeutic targets. However, experimental validation is necessary to confirm these predictions. Our study comes into contributions to the development of personalized target therapy for pancreatic cancer patients.展开更多
Metabolism is a general term for a series of ordered chemical reactions in an organism used to maintain life,mainly divided into anabolic and catabolic metabolism.Nucleic acid therapy can not only precisely up-regulat...Metabolism is a general term for a series of ordered chemical reactions in an organism used to maintain life,mainly divided into anabolic and catabolic metabolism.Nucleic acid therapy can not only precisely up-regulate and down-regulate the expression of target genes but also correct mutated disease-causing genes,which demonstrates irreplaceable and outstanding advantages in the treatment of metabolismrelated diseases and has been applied to the clinical treatment of metabolism-related diseases.In this review,we introduce the structures of several major nucleic acid drugs and the mechanism of nucleic acid therapy.Subsequently,we describe the mechanisms of various biomolecular and tissue metabolisms and the etiology of metabolic disorders,classified according to metabolic substrates.We analyze the signal pathways and potential targets affecting the metabolism of each substrate and describe the nucleic acid drugs applied to these targets and their delivery technologies.This review aims to provide new ideas and targets for treating these diseases by investigating the role played by metabolism in developing diseases and providing guidance for the selection and design of nucleic acid drugs.展开更多
Alzheimer＇s disease（AD） is the most common form of dementia in the older population, however, the precise cause of the disease is unknown. The neuropathology is characterized by the presence of aggregates formed by...Alzheimer＇s disease（AD） is the most common form of dementia in the older population, however, the precise cause of the disease is unknown. The neuropathology is characterized by the presence of aggregates formed by amyloid-β（Aβ） peptide and phosphorylated tau; which is accompanied by progressive impairment of memory. Diverse signaling pathways are linked to AD, and among these the Wnt signaling pathway is becoming increasingly relevant, since it plays essential roles in the adult brain. Initially, Wnt signaling activation was proposed as a neuroprotective mechanism against Aβ toxicity. Later, it was reported that it participates in tau phosphorylation and processes of learning and memory. Interestingly, in the last years we demonstrated that Wnt signaling is fundamental in amyloid precursor protein（APP） processing and that Wnt dysfunction results in Aβ production and aggregation in vitro. Recent in vivo studies reported that loss of canonical Wnt signaling exacerbates amyloid deposition in a transgenic（Tg） mouse model of AD. Finally, we showed that inhibition of Wnt signaling in a Tg mouse previously at the appearance of AD signs, resulted in memory loss, tau phosphorylation and Aβ formation and aggregation; indicating that Wnt dysfunction accelerated the onset of AD. More importantly, Wnt signaling loss promoted cognitive impairment, tau phosphorylation and Aβ1–42 production in the hippocampus of wild-type（WT） mice, contributing to the development of an Alzheimer＇s-like neurophatology. Therefore, in this review we highlight the importance of Wnt/β-catenin signaling dysfunction in the onset of AD and propose that the loss of canonical Wnt signaling is a triggering factor of AD.展开更多
Axon regeneration is crucial for recovery from neurological diseases. Numerous studies have identified several genes, microRNAs （miRNAs）, and transcription factors （TFs） that influence axon regeneration. However, ...Axon regeneration is crucial for recovery from neurological diseases. Numerous studies have identified several genes, microRNAs （miRNAs）, and transcription factors （TFs） that influence axon regeneration. However, the regulatory networks involved have not been fully elucidated. In the present study, we analyzed a regulatory network of 51 miRNAs, 27 TFs, and 59 target genes, which is involved in axon regeneration. We identified 359 pairs of feed- forward loops （FFLs）, seven important genes （Naplll, Arhgef12, Sema6d, Akt3, Trim2, Rabllfip2, and Rps6ka3）, six important miRNAs （hsa-miR-204-5p, hsa-miR-124-3p, hsa-miR-26a-5p, hsa-miR-16-5p, hsa-miR-17-5p, and hsa- miR-15b-5p）, and eight important TFs （Smada2, Flil, Wtl, Sp6, Sp3, Smad4, Smad5, and Crebl）, which appear to play an important role in axon regeneration. Functional enrichment analysis revealed that axon-associated genes are involved mainly in the regulation of cellular component organization, axonogenesis, and cell morphogenesis during neuronal differentiation. However, these findings need to be validated by further studies.展开更多
The long-arm and short-arm genes of fibroin light chain (L-chain) of silkworm, Bombyx Mori L., and the gene of human acidic fibroblast growth factor were cloned respectively and subsequently inserted into a transfer v...The long-arm and short-arm genes of fibroin light chain (L-chain) of silkworm, Bombyx Mori L., and the gene of human acidic fibroblast growth factor were cloned respectively and subsequently inserted into a transfer vector pVL 1392 used as a tool to target the L-chain region of the silkworm genome. Genomic DNA from their offsprings was extracted and the expected targeting was detected using polymerase chain reaction and DNA sequencing, as well as protein analysis. The results showed that positive events occurred and that the FGF gene was integrated into the L-chain locus through homologous recombination.展开更多
Autophagy is a widely conserved intracellular process for degradation and recycling of proteins, organelles and cytoplasm in eukaryotic organisms and is now emerging as an important process in tbliar infection by many...Autophagy is a widely conserved intracellular process for degradation and recycling of proteins, organelles and cytoplasm in eukaryotic organisms and is now emerging as an important process in tbliar infection by many plant pathogenic fungi. However, the role of autophagy in soil-borne fungal physiology and infection biology is poorly understood. Here, we report the establishment of an Agro- bacterium tumefaciens-mediated transformation （ATMT） system and its application to investigate two autophagy genes, VdATG8 and VdATG12, by means of targeted gene replacement and complementation. Transformation of a cotton-infecting Verticillium dahliae strain Vd8 with a novel binary vector pCOM led to the production of 384 geneticin-resistant translbnnants per 1 × 10^4 conidia. V. dahliae mutants lacking either VdATG8 or VdATGI2 exhibited reduced conidiation and impaired aerial hyphae production. Disease development on Arabidopsis plants was slightly delayed when inoculated with VdATG8 or VdATG12 gene deletion mutants, compared with the wild- type and gene complemented strains. Surprisingly, in vitro inoculation with unimpaired roots revealed that the abilities of root invasion were not affected in gene deletion mutants. These results indicate that autophagy is necessary for aerial hyphae development and plant colonization but not for root infection in E dahliae.展开更多
AIM To evaluate associations between mi RNA target genes IL12B,INSR,CCND1 and IL10 polymorphisms and gastric cancer(GC)in European population.METHODS Gene polymorphisms were analyzed in 508 controls and474 GC patients...AIM To evaluate associations between mi RNA target genes IL12B,INSR,CCND1 and IL10 polymorphisms and gastric cancer(GC)in European population.METHODS Gene polymorphisms were analyzed in 508 controls and474 GC patients from 3 tertiary centers in Germany,Lithuania and Latvia.Controls were patients from the out-patient departments,who were referred for upper endoscopy because of dyspeptic symptoms and had no history of previous malignancy.Gastric cancer(GC)patients had histopathological verification of gastric adenocarcinoma.Genomic DNA was extracted using salting out method from peripheral blood mononuclear cells.IL12B T>G(rs1368439),INSR T>C(rs1051690),CCND1 A>C(rs7177)and IL10 T>C(rs3024498)SNPs were genotyped by the real-time polymerase chain reaction.Associations between gene polymorphism and GC were evaluated using multiple logistic regression analysis with adjustment for sex,age and country of birth.RESULTS We observed similar distribution of genotypes and allelic frequencies of all polymorphisms between GC patients and controls except of INSR rs1051690.The frequency of the T allele of INSR gene was significantly higher in GC patients than in controls(23.26%and 19.19%respectively,P=0.028).CT genotype was also more prevalent in patients compared to control group(38.48%and 30.12%respectively,P<0.021).Logistic regression analysis revealed that only one polymorphism(rs1051690 in INSR gene)was associated with increased risk of GC.Carriers of CT genotype had higher odds of GC when compared to CC genotype(OR=1.45,95%PI:1.08-1.95,P=0.01).Similar association was observed in a dominant model for INSR gene,where comparison of TT+CT vs CC genotypes showed an increased risk of GC(OR=1.44,95%PI:1.08-1.90,P=0.01).Other analyzed SNPs were not associated with the presence of GC.CONCLUSION INSR rs1051690 SNP is associated with increased risk of GC,while polymorphisms in IL12B,CCND1 and IL10genes are not linked with the presence of GC.展开更多
High grain protein content(GPC) reduces rice eating and cooking quality(ECQ). We generated OsAAP6 and OsAAP10 knockout mutants in three high-yielding japonica varieties and one japonica line using the CRISPR/Cas9 syst...High grain protein content(GPC) reduces rice eating and cooking quality(ECQ). We generated OsAAP6 and OsAAP10 knockout mutants in three high-yielding japonica varieties and one japonica line using the CRISPR/Cas9 system. Mutation efficiency varied with genetic background in the T_0 generation, and GPC in the T_1 generation decreased significantly,owing mainly to a reduction in glutelin content. Amylose content was down-regulated significantly in some Osaap6 and all Osaap10 mutants. The increased taste value of these mutants was supported by Rapid Visco Analysis(RVA) profiles, which showed higher peak viscosity and breakdown viscosity and lower setback viscosity than the wild type. There were no significant deficiencies in agronomic traits of the mutants. Targeted mutagenesis of OsAAP6 and OsAAP10, especially OsAAP10, using the CRISPR/Cas9 system can rapidly reduce GPC and improve ECQ of rice, providing a new strategy for the breeding cultivars with desired ECQ.展开更多
Targeted genome modifications with the Cas9/gRNA system derived from the clustered regularly interspaced short palin- dromic repeat/CRISPR-associated （CRISPR/Cas） system have been successfully used in cultured human...Targeted genome modifications with the Cas9/gRNA system derived from the clustered regularly interspaced short palin- dromic repeat/CRISPR-associated （CRISPR/Cas） system have been successfully used in cultured human cells as well as in most model organisms, including zebrafish （Danio rerio）, mouse, and fruit fly （Chang et al., 2013; Cong et al., 2013; Gratz et al., 2013; Hwang et al., 2013; Jao et al., 2013; Shen et al., 2013; Wei et al., 2013）. Its application in zebrafish is particu- larly attractive due to the ease of handling this organism and the simple application of this method by direct injection of Cas9/ gRNA. However, the information about its specificity in this organism is very limited and needs further evaluation. In addition, it is conceivable that a Cas9 mRNA optimized for zebrafish codon preference could enhance its activity.展开更多
Penicillium digitatum is the most important pathogen of postharvest citrus. Gene targeting can be done in P. digitatum using homologous recombination via Agrobacterium tumefaciens mediated transformation （ATMT）, but...Penicillium digitatum is the most important pathogen of postharvest citrus. Gene targeting can be done in P. digitatum using homologous recombination via Agrobacterium tumefaciens mediated transformation （ATMT）, but the frequencies are often very low. In the present study, we replaced the Ku80 homolog （a gene of the non-homologous end-joining （NHEJ） pathway） with the hygromycin resistance cassette （hph） by ATMT. No significant change in vegetative growth, conidiation, or pathogenicity was observed in KuSO-deficient strain （△dKuSO） of P. digitatum. However, using △pdKuSO as a targeting strain, the gene-targeting frequencies for both genes PdbrlA and PdmpkA were significantly increased. These results suggest that Ku80 plays an important role in homologous inte- gration and the created △PdKuSO strain would be a good candidate for rapid gene function analysis in P. digitatum.展开更多
CRISPR （clustered regularly interspaced short palindromic repeats）-Cas9-based genome editing has revolutionized func- tional genomics in many biological research fields. The specificity and potency of CR1SPR-Cas9 ge...CRISPR （clustered regularly interspaced short palindromic repeats）-Cas9-based genome editing has revolutionized func- tional genomics in many biological research fields. The specificity and potency of CR1SPR-Cas9 genome editing make it ideal for investigating the function of genes in vivo （Hsu et al., 2014）. Gene duplication is a key driver of evolu- tionary novelty （Taylor and Raes, 2004）. However, duplicated genes with near-identical sequences and functional redun- dancy have posed challenges for genetic analysis （Woollard, 2005）. The functions of duplicated genes can be assessed by simultaneous knockdown using homology-based methods such as RNA interference （RNAi） （Tischler et al., 2006）, Generation of double or triple mutants is an alternative way to assess functional redundancy of duplicated genes, However, generation of such compound mutants by forward or reverse genetic methods is time consuming.展开更多
Background:Screening key target genes for pulmonary arterial hypertension(PAH)based on bioinformatics to provide a reference for the clinical development of drugs to cure PAH.Methods:The keyword“pulmonary arterial hy...Background:Screening key target genes for pulmonary arterial hypertension(PAH)based on bioinformatics to provide a reference for the clinical development of drugs to cure PAH.Methods:The keyword“pulmonary arterial hypertension”was used to search related genes in the National Center for Biotechnology Information database(NCBI).The obtained genes data was input to the database of Database for Annotation,Visualization and Integrated Discovery(DAVID)(Version 6.8)to collect relevant information about pathways and genes.And the data of genes were enriched in 37 pathways and genes with occurrence frequency≥10 were respectively imported into the String database to construct protein-protein interaction(PPI)network diagrams,and the two network diagrams were compared.Results:VEGFA,MAPK1,MAPK3,IL6,JUN and TNF were among the highest-ranked genes in two network diagrams.Conclusion:The pathogenesis of PAH is associated with multiple pathways such as the TGF-βsignaling pathway,PI3K-Akt signaling pathway,MAPK signaling pathway,HIF-1 signaling pathway and so on.The study of VEGFA,MAPK1,MAPK3,IL6,JUN and TNF are closely related to PAH is necessary for us to study further.Through gene interaction network and pathway analysis of disease-associated genes,which will help us to screen the critical target genes of PAH and provide a reference for clinical development of effective drugs for PAH.展开更多
BACKGROUND Approximately half of all new cases of gastric cancer(GC)and related deaths occur in China.More than 80%of patients with GC are diagnosed at an advanced stage,which results in poor prognosis.Although HER2-d...BACKGROUND Approximately half of all new cases of gastric cancer(GC)and related deaths occur in China.More than 80%of patients with GC are diagnosed at an advanced stage,which results in poor prognosis.Although HER2-directed therapy and immune checkpoint inhibitors have been somewhat successful,new drugs are still needed for the treatment of GC.Notably,several gene fusion-targeted drugs have been approved by the United States Food and Drug Administration for solid tumors,including GC,such as larotrectinib for NTRK fusion-positive cancers and zenocutuzumab for NRG1 fusion-positive cancers.However,gene fusions involving targetable genes have not been well characterized in Chinese patients with GC.AIM To identify the profile of fusions involving targetable genes in Chinese patients with GC using clinical specimens and determine the distribution of patients with gene fusion variants among the molecular subtypes of GC.METHODS We retrospectively analyzed gene fusion events in tumor tissue samples from 954 Chinese patients with GC.Clinicopathological characteristics were obtained from their medical records.Genetic alterations,such as single nucleotide variants,indels,amplifications,and gene fusions,were identified using a targeted sequencing panel containing 825 genes.Fusions were validated by fluorescence in situ hybridization(FISH)using break-apart probes.The microsatellite instability(MSI)status was evaluated using MSIsensor from the targeted sequencing panel data.Tumor mutational burden(TMB)was calculated using the total number of nonsynonymous mutations divided by the total genomic targeted region.Chi-square analysis was used to determine the enrichment of gene fusions associated with the molecular subtypes of GC.RESULTS We found that 1.68%(16/954)of patients harbored 20 fusion events involving targetable genes.RARA fusions(n=5)were the most common,followed by FGFR2,BRAF,MET,FGFR3,RET,ALK,EGFR,NTRK2,and NRG1 fusions.Two of the RARA fusions,EML4-ALK(E6:E20)and EGFRSEPTIN14(E7:E10),have been identified in other tumors but not in GC.Surprisingly,18 gene fusion events were previously not reported in any cancer types.Twelve of the eighteen novel gene fusions included complete exons encoding functional domains of targetable genes,such as the tyrosine kinase domain of receptor tyrosine kinases and the DNA-and ligand-binding domains of RARA.Consistent with the results of detection using the targeted sequencing fusion panel,the results of FISH(fluorescence in situ hybridization)confirmed the rearrangement of FGFR2 and BRAF in tumors from patients 04 and 09,respectively.Genetic analysis indicated that the fusion genes were significantly enriched in patients with ERBB2 amplification(P=0.02);however,there were no significant differences between fusion-positive and fusion-negative patients in age,sex,MSI status,and TMB.CONCLUSION We characterized the landscape of fusions involving targetable genes in a Chinese GC cohort and found that 1.68%of patients with GC harbor potential targetable gene fusions,which were enriched in patients with ERBB2 amplification.Gene fusion detection may provide a potential treatment strategy for patients with GC with disease progression following standard therapy.展开更多
The classic method for gene knockout (KO) is based on homologous recombination (HR) and embryonic stem cell technique (Gerlai,1996).Actually,the procedure of homologous replacement is complicated and time consuming,al...The classic method for gene knockout (KO) is based on homologous recombination (HR) and embryonic stem cell technique (Gerlai,1996).Actually,the procedure of homologous replacement is complicated and time consuming,although it has been popular during the past decades.Recent years,genome editing which can cause DNA sequence-specific mutations in the genomes of cellular展开更多
Objective:To investigate the core target genes of miR-29b-3p,and analyze the clinical significance of the core target genes in glioma.Methods:Bioinformatics analysis was used to predict and screen the target genes of ...Objective:To investigate the core target genes of miR-29b-3p,and analyze the clinical significance of the core target genes in glioma.Methods:Bioinformatics analysis was used to predict and screen the target genes of miR-29b-3p.STRING and Cytoscape software were used to analyze the protein-protein interaction(PPI)of target genes.the differences expression and survival prognosis in glioma were analyzed by GEPIA and CGGA.Independent prognostic factors analyzed by univariate and multivariate Cox proportional hazards regression model.Results:22 target genes of miR-29b-3p were predicted using LinkedOmics,miRDB,miRTarBase,TargetScan,and starbase databases.Through the construction of the PPI network,genes out of the network were removed,and a total of 16 genes were screened for further study of their clinical significance.Based on analysis of GEPIA and CGGA databases,COL2A1,DNMT3A,and DNMT3B were excluded.Through further analysis of the univariate and multivariate Cox proportional hazard regression model,finally identified three core target genes:SERPINH1,LOXL2,CDK6.Conclusion:Bioinformatics analysis showed that miR-29b-3p targeted three core genes such as SERPINH1,LOXL2,and CDK6 in glioma.The expression of these genes was different between brain normal tissues and gliomas,between different grades of tumor,IDH mutation status and 1p/19q codeletion status.Its high expression had adverse effects on overall survival and recurrence-free survival.These core target genes can be used as an independent prognostic factor.展开更多
Haemophilus parasuis is one kind of constant bacteria in porcine upper respiratory tract, and it can cause multiple serositis, arthritis and other diseases under certain conditions. Due to lack of efficient genetic op...Haemophilus parasuis is one kind of constant bacteria in porcine upper respiratory tract, and it can cause multiple serositis, arthritis and other diseases under certain conditions. Due to lack of efficient genetic operating system, its pathogenic mechanism is not very clear. Ligation with DNA ligase and fusion PCR were used to construct targeting hhdA gene of Haemophilus parasuis, respectively. The fidelity, application scope, operation and conditions of the constructed fusion fragments were compared. The results showed that construction with DNA ligase was more mature technology as manifested by more stable conditions and more extensive application. The fusion PCR method had high fidelity and simple operation, and the transformation rate was 9.5 times as high as that of ligation with DNA ligase. For this reason, this method was more suitable for construction of multi-fragment targeting genes. The study lays a foundation for establishing an efficient operating system of targeting gene of Haemophilus parasuis in the future.展开更多
[ Objective] This study aimed to screen target genes regulated by heat shock factor AtHsfAla in Arabidopsis thaliana. [ Method] Using AtHsfAla-in- serted mutant athsfala （SALK-068042） and wild-type A. thaliana seedl...[ Objective] This study aimed to screen target genes regulated by heat shock factor AtHsfAla in Arabidopsis thaliana. [ Method] Using AtHsfAla-in- serted mutant athsfala （SALK-068042） and wild-type A. thaliana seedlings as experimental materials, target genes regulated by heat shock factor AtHsfAla were screened by microarray assay. Differentially expressed genes were screened by multiple method. Specific functions of differentially expressed genes were analyzed by gene ontology （GO） analysis. Signal transduction pathways, in which differentia｜ly expressed genes were involved, were analyzed by pathway analysis. Gene-gene interaction network was constructed by Signal-Net. [ Result] A total of 3 672 differentially expressed genes were screened out. Up-regulated differentially expressed genes were involved in 198 functions and 7 signal transduction pathways; down-regulated differentially expressed genes were involved in 94 functions and 10 signal transduction pathways. In the signal transduction network, it was found that cwlNV4 and HXK3 had relatively high ability of mediation; AT1 G14240 and cwlNV4 ex- hibited the most interactions with other genes, which were located in key positions throughout the gene-gene interaction network. [ Conclusion] Heat shock factor AtHsfAla regulates a large number of target genes in A. thaliana.展开更多
Gene targeting technology is an important means to investigate gene functions, but its efficiency of gene targeting is very low, especially for somatic cell targeting. Artificially induced double-strand breaks （DSB）...Gene targeting technology is an important means to investigate gene functions, but its efficiency of gene targeting is very low, especially for somatic cell targeting. Artificially induced double-strand breaks （DSB） and triplex forming oligonucleotide （TFO） are currently developed methods to improve the targeting efficiency. This paper summarized the basic principles, design ideas and application in gene targeting efficiency improvement of these two methods, analyzed and com- pared their characteristics, and finally proposed prospects for their future development.展开更多
基金supported by the National Key Research and Development Program of China(2022YFF1001502)Jiangsu Collaborative Innovation Center for Modern Crop Production(JCIC-MCP)Major Projects of Technological Innovation 2030(2023ZD04037).
文摘Soybean mosaic virus(SMV)poses a substantial threat to the yield and quality of soybean(Glycine max(L.)Merr.),leading to significant economic losses in soybean production.However,the mining of SMVresistance loci and the exploration of the underlying disease resistance mechanisms remain relatively limited.MicroRNAs(miRNAs)are a class of post-transcriptional regulators that play a pivotal role in modulating plant growth,development and responding to various stresses.In this study,we demonstrated the function of the “miR398c/d-GmCSDs”module between soybean resistant and susceptible varieties,focusing on its differential regulatory roles in SMV infection.Specifically,SMV infection downregulated gma-miR398c/d expression in the resistant variety(Qihuang 1,QH),while upregulated them in the susceptible variety(Nannong 1138-2,NN).Transient expression assay in N.benthamiana confirmed that gma-miR398c/d can target six superoxide dismutase(SOD)family genes,which responded to SMV infection in both varieties.Stable overexpression of Gma-MIR398c/d in soybean or inhibition of the corresponding target genes’expression via Bean pod mottle virus(BPMV)-induced gene silencing(VIGS)led to reduced H_(2)O_(2)content and thereby promoted SMV infection.Conversely,plants overexpressing the target genes exhibited the opposite phenotypes.The functions of gma-miR398c/d and their target genes were further validated in N.benthamiana through transient co-expression with SMV infectious clone(pSC7-GFP),indicating that gma-miR398c/d negatively regulated soybean resistance to SMV,while the target genes positively contributed to disease resistance.Collectively,our findings provide novel insights into the regulatory mechanisms underlying soybean resistance to SMV.
文摘Elevated lipoprotein(a)[Lp(a)]is a major independent risk factor for atheroscle-rotic cardiovascular disease(ASCVD),with limited response to traditional lipid-lowering therapies.Lepodisiran,a novel N-acetylgalactosamine-conjugated small interfering RNA,targets hepatic LPA message RNA to reduce apolipoprotein(a)production.Early-phase trials demonstrated>90%sustained Lp(a)reduction with excellent safety and tolerability.The phase 2 ALPACA trial confirmed dura-ble effects lasting up to one year after biannual dosing.Compared to other thera-pies,lepodisiran offers longer duration,high efficacy,and minimal side effects.Ongoing phase 3 studies aim to determine its impact on cardiovascular outcomes,potentially establishing a new standard in precise ASCVD risk management.
文摘Objective: The mortality and morbidity rates associated with pancreatic cancer (PaCa) are extremely high. Various studies have demonstrated that pancreatic cancer will be the fourth cancer-related death by 2030, raising more concern for scholars to find effective methods to prevent and treat in order to improve the pancreatic cancer outcome. Using bioinformatic analysis, this study aims to pinpoint key genes that could impact PaCa patients’ prognosis and could be used as therapeutic targets. Methods: The TCGA and GEO datasets were integratively analyzed to identify prognosis-related differentially expressed genes. Next, the STRING database was used to develop PPI networks, and the MCODE and CytoNCA Cytoscape in Cytoscape were used to screen for critical genes. Through CytoNCA, three kinds of topology analysis were considered (degree, betweenness, and eigenvector). Essential genes were confirmed as potential target treatment through Go function and pathways enrichment analysis, a developed predictive risk model based on multivariate analysis, and the establishment of nomograms using the clinical information. Results: Overall, the GSE183795 and TCGA datasets associated 1311 and 2244 genes with pancreatic cancer prognosis, respectively. We identified 132 genes that were present in both datasets. The PPI network analysis using, the centrality analysis approach with the CytoNCA plug-in, showed that CDK2, PLK1, CCNB1, and TOP2A ranked in the top 5% across all three metrics. The independent analysis of a risk model revealed that the four key genes had a Hazard Ratio (HR) > 1. The monogram showed the predictive risk model and individual patient survival predictions were accurate. The results indicate that the effect of the selected vital genes was significant and that they could be used as biomarkers to predict a patient’s outcome and as possible target therapy in patients with pancreatic cancer. GO function and pathway analysis demonstrated that crucial genes might affect the P53 signaling pathway and FoxO signaling pathway, through which Meiotic nuclear division and cell cycle may have a significant function in essential genes affecting the outcome of patients who have pancreatic cancer. Conclusions: This study suggests that CDK2, CCNB1, PLK1 and TOP2A are four key genes that have a significant influence on PaCa migration and proliferation. CDK2, CCNB1, PLK1, and TOP2A can be used as potential PaCa prognostic biomarkers and therapeutic targets. However, experimental validation is necessary to confirm these predictions. Our study comes into contributions to the development of personalized target therapy for pancreatic cancer patients.
基金financially supported by the National Natural Science Foundation of China(Nos.32225029,22205240,52073287,22075289,82071552 and 22376006)National Key R&D Program of China(No.2023YFC2605003)。
文摘Metabolism is a general term for a series of ordered chemical reactions in an organism used to maintain life,mainly divided into anabolic and catabolic metabolism.Nucleic acid therapy can not only precisely up-regulate and down-regulate the expression of target genes but also correct mutated disease-causing genes,which demonstrates irreplaceable and outstanding advantages in the treatment of metabolismrelated diseases and has been applied to the clinical treatment of metabolism-related diseases.In this review,we introduce the structures of several major nucleic acid drugs and the mechanism of nucleic acid therapy.Subsequently,we describe the mechanisms of various biomolecular and tissue metabolisms and the etiology of metabolic disorders,classified according to metabolic substrates.We analyze the signal pathways and potential targets affecting the metabolism of each substrate and describe the nucleic acid drugs applied to these targets and their delivery technologies.This review aims to provide new ideas and targets for treating these diseases by investigating the role played by metabolism in developing diseases and providing guidance for the selection and design of nucleic acid drugs.
基金supported by grants PFB (Basal Financing Program) 12/2007 from the Basal Centre for Excellence in Science and Technology and FONDECYT,No.1120156(to NCI)a pre-doctoral fellowship from the National Commission of Science and Technology of Chile(CONICYT)(to CTR)
文摘Alzheimer＇s disease（AD） is the most common form of dementia in the older population, however, the precise cause of the disease is unknown. The neuropathology is characterized by the presence of aggregates formed by amyloid-β（Aβ） peptide and phosphorylated tau; which is accompanied by progressive impairment of memory. Diverse signaling pathways are linked to AD, and among these the Wnt signaling pathway is becoming increasingly relevant, since it plays essential roles in the adult brain. Initially, Wnt signaling activation was proposed as a neuroprotective mechanism against Aβ toxicity. Later, it was reported that it participates in tau phosphorylation and processes of learning and memory. Interestingly, in the last years we demonstrated that Wnt signaling is fundamental in amyloid precursor protein（APP） processing and that Wnt dysfunction results in Aβ production and aggregation in vitro. Recent in vivo studies reported that loss of canonical Wnt signaling exacerbates amyloid deposition in a transgenic（Tg） mouse model of AD. Finally, we showed that inhibition of Wnt signaling in a Tg mouse previously at the appearance of AD signs, resulted in memory loss, tau phosphorylation and Aβ formation and aggregation; indicating that Wnt dysfunction accelerated the onset of AD. More importantly, Wnt signaling loss promoted cognitive impairment, tau phosphorylation and Aβ1–42 production in the hippocampus of wild-type（WT） mice, contributing to the development of an Alzheimer＇s-like neurophatology. Therefore, in this review we highlight the importance of Wnt/β-catenin signaling dysfunction in the onset of AD and propose that the loss of canonical Wnt signaling is a triggering factor of AD.
基金Project supported by the Key Project of Hebei North University(No.120177)the Science and Technology Bureau Research Development Plan of Zhangjiakou City in Hebei(No.0911021D-4)China
文摘Axon regeneration is crucial for recovery from neurological diseases. Numerous studies have identified several genes, microRNAs （miRNAs）, and transcription factors （TFs） that influence axon regeneration. However, the regulatory networks involved have not been fully elucidated. In the present study, we analyzed a regulatory network of 51 miRNAs, 27 TFs, and 59 target genes, which is involved in axon regeneration. We identified 359 pairs of feed- forward loops （FFLs）, seven important genes （Naplll, Arhgef12, Sema6d, Akt3, Trim2, Rabllfip2, and Rps6ka3）, six important miRNAs （hsa-miR-204-5p, hsa-miR-124-3p, hsa-miR-26a-5p, hsa-miR-16-5p, hsa-miR-17-5p, and hsa- miR-15b-5p）, and eight important TFs （Smada2, Flil, Wtl, Sp6, Sp3, Smad4, Smad5, and Crebl）, which appear to play an important role in axon regeneration. Functional enrichment analysis revealed that axon-associated genes are involved mainly in the regulation of cellular component organization, axonogenesis, and cell morphogenesis during neuronal differentiation. However, these findings need to be validated by further studies.
基金Project supported by the ScientificResearch Foundation forReturned Overseas Chinese Scholars,Education Ministry of Chinaand the Natural Science Foundation of Zhejiang Province (No.301306), China
文摘The long-arm and short-arm genes of fibroin light chain (L-chain) of silkworm, Bombyx Mori L., and the gene of human acidic fibroblast growth factor were cloned respectively and subsequently inserted into a transfer vector pVL 1392 used as a tool to target the L-chain region of the silkworm genome. Genomic DNA from their offsprings was extracted and the expected targeting was detected using polymerase chain reaction and DNA sequencing, as well as protein analysis. The results showed that positive events occurred and that the FGF gene was integrated into the L-chain locus through homologous recombination.
基金supported by the grants from the State Key Basic Research and Development Plan of China(No. 2011CB109300)the National Natural Science Foundation of China(No.31171590)+2 种基金Jiangsu Province Natural Science Foundation(Nos.BK2010065 and BE2012329)the Specialized Research Fund for the Doctoral Program of Higher Education of China(No.20090097110010)the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘Autophagy is a widely conserved intracellular process for degradation and recycling of proteins, organelles and cytoplasm in eukaryotic organisms and is now emerging as an important process in tbliar infection by many plant pathogenic fungi. However, the role of autophagy in soil-borne fungal physiology and infection biology is poorly understood. Here, we report the establishment of an Agro- bacterium tumefaciens-mediated transformation （ATMT） system and its application to investigate two autophagy genes, VdATG8 and VdATG12, by means of targeted gene replacement and complementation. Transformation of a cotton-infecting Verticillium dahliae strain Vd8 with a novel binary vector pCOM led to the production of 384 geneticin-resistant translbnnants per 1 × 10^4 conidia. V. dahliae mutants lacking either VdATG8 or VdATGI2 exhibited reduced conidiation and impaired aerial hyphae production. Disease development on Arabidopsis plants was slightly delayed when inoculated with VdATG8 or VdATG12 gene deletion mutants, compared with the wild- type and gene complemented strains. Surprisingly, in vitro inoculation with unimpaired roots revealed that the abilities of root invasion were not affected in gene deletion mutants. These results indicate that autophagy is necessary for aerial hyphae development and plant colonization but not for root infection in E dahliae.
基金Supported by Lithuanian Research Council Grant,No.MIP-14418
文摘AIM To evaluate associations between mi RNA target genes IL12B,INSR,CCND1 and IL10 polymorphisms and gastric cancer(GC)in European population.METHODS Gene polymorphisms were analyzed in 508 controls and474 GC patients from 3 tertiary centers in Germany,Lithuania and Latvia.Controls were patients from the out-patient departments,who were referred for upper endoscopy because of dyspeptic symptoms and had no history of previous malignancy.Gastric cancer(GC)patients had histopathological verification of gastric adenocarcinoma.Genomic DNA was extracted using salting out method from peripheral blood mononuclear cells.IL12B T>G(rs1368439),INSR T>C(rs1051690),CCND1 A>C(rs7177)and IL10 T>C(rs3024498)SNPs were genotyped by the real-time polymerase chain reaction.Associations between gene polymorphism and GC were evaluated using multiple logistic regression analysis with adjustment for sex,age and country of birth.RESULTS We observed similar distribution of genotypes and allelic frequencies of all polymorphisms between GC patients and controls except of INSR rs1051690.The frequency of the T allele of INSR gene was significantly higher in GC patients than in controls(23.26%and 19.19%respectively,P=0.028).CT genotype was also more prevalent in patients compared to control group(38.48%and 30.12%respectively,P<0.021).Logistic regression analysis revealed that only one polymorphism(rs1051690 in INSR gene)was associated with increased risk of GC.Carriers of CT genotype had higher odds of GC when compared to CC genotype(OR=1.45,95%PI:1.08-1.95,P=0.01).Similar association was observed in a dominant model for INSR gene,where comparison of TT+CT vs CC genotypes showed an increased risk of GC(OR=1.44,95%PI:1.08-1.90,P=0.01).Other analyzed SNPs were not associated with the presence of GC.CONCLUSION INSR rs1051690 SNP is associated with increased risk of GC,while polymorphisms in IL12B,CCND1 and IL10genes are not linked with the presence of GC.
基金financially supported by National Key Research and Development Program of China(2016YFD0100501)the National Natural Science Foundation of China(31871241,31371233)+3 种基金the Natural Science Foundation of Jiangsu Province(BE2017345,PZCZ201702,BE2018351)the Research and Innovation Program of Postgraduate in Jiangsu Province(KYCX17_1886)the Priority Academic Program Development of Jiangsu Higher Education Institutionsthe Yangzhou University International Academic Exchange Fund。
文摘High grain protein content(GPC) reduces rice eating and cooking quality(ECQ). We generated OsAAP6 and OsAAP10 knockout mutants in three high-yielding japonica varieties and one japonica line using the CRISPR/Cas9 system. Mutation efficiency varied with genetic background in the T_0 generation, and GPC in the T_1 generation decreased significantly,owing mainly to a reduction in glutelin content. Amylose content was down-regulated significantly in some Osaap6 and all Osaap10 mutants. The increased taste value of these mutants was supported by Rapid Visco Analysis(RVA) profiles, which showed higher peak viscosity and breakdown viscosity and lower setback viscosity than the wild type. There were no significant deficiencies in agronomic traits of the mutants. Targeted mutagenesis of OsAAP6 and OsAAP10, especially OsAAP10, using the CRISPR/Cas9 system can rapidly reduce GPC and improve ECQ of rice, providing a new strategy for the breeding cultivars with desired ECQ.
基金partially supported by the National Natural Science Foundation of China (No. 31110103904)the National Program on Key Basic Research Project (973 Program) of the Ministry of Science and Technology of China (Nos. 2011CBA01000 and 2012CB945101)
文摘Targeted genome modifications with the Cas9/gRNA system derived from the clustered regularly interspaced short palin- dromic repeat/CRISPR-associated （CRISPR/Cas） system have been successfully used in cultured human cells as well as in most model organisms, including zebrafish （Danio rerio）, mouse, and fruit fly （Chang et al., 2013; Cong et al., 2013; Gratz et al., 2013; Hwang et al., 2013; Jao et al., 2013; Shen et al., 2013; Wei et al., 2013）. Its application in zebrafish is particu- larly attractive due to the ease of handling this organism and the simple application of this method by direct injection of Cas9/ gRNA. However, the information about its specificity in this organism is very limited and needs further evaluation. In addition, it is conceivable that a Cas9 mRNA optimized for zebrafish codon preference could enhance its activity.
基金supported by the National Natural Science Foundation of China(Nos.31371961 and 31071649)the China Agriculture Research System(No.CARS-27)the Special Fund for Agro-scientific Research in the Public Interest(No.201203034),China
文摘Penicillium digitatum is the most important pathogen of postharvest citrus. Gene targeting can be done in P. digitatum using homologous recombination via Agrobacterium tumefaciens mediated transformation （ATMT）, but the frequencies are often very low. In the present study, we replaced the Ku80 homolog （a gene of the non-homologous end-joining （NHEJ） pathway） with the hygromycin resistance cassette （hph） by ATMT. No significant change in vegetative growth, conidiation, or pathogenicity was observed in KuSO-deficient strain （△dKuSO） of P. digitatum. However, using △pdKuSO as a targeting strain, the gene-targeting frequencies for both genes PdbrlA and PdmpkA were significantly increased. These results suggest that Ku80 plays an important role in homologous inte- gration and the created △PdKuSO strain would be a good candidate for rapid gene function analysis in P. digitatum.
基金supported by National Institutes of Health(NIH grant R01GM054657)to A.D.C
文摘CRISPR （clustered regularly interspaced short palindromic repeats）-Cas9-based genome editing has revolutionized func- tional genomics in many biological research fields. The specificity and potency of CR1SPR-Cas9 genome editing make it ideal for investigating the function of genes in vivo （Hsu et al., 2014）. Gene duplication is a key driver of evolu- tionary novelty （Taylor and Raes, 2004）. However, duplicated genes with near-identical sequences and functional redun- dancy have posed challenges for genetic analysis （Woollard, 2005）. The functions of duplicated genes can be assessed by simultaneous knockdown using homology-based methods such as RNA interference （RNAi） （Tischler et al., 2006）, Generation of double or triple mutants is an alternative way to assess functional redundancy of duplicated genes, However, generation of such compound mutants by forward or reverse genetic methods is time consuming.
文摘Background:Screening key target genes for pulmonary arterial hypertension(PAH)based on bioinformatics to provide a reference for the clinical development of drugs to cure PAH.Methods:The keyword“pulmonary arterial hypertension”was used to search related genes in the National Center for Biotechnology Information database(NCBI).The obtained genes data was input to the database of Database for Annotation,Visualization and Integrated Discovery(DAVID)(Version 6.8)to collect relevant information about pathways and genes.And the data of genes were enriched in 37 pathways and genes with occurrence frequency≥10 were respectively imported into the String database to construct protein-protein interaction(PPI)network diagrams,and the two network diagrams were compared.Results:VEGFA,MAPK1,MAPK3,IL6,JUN and TNF were among the highest-ranked genes in two network diagrams.Conclusion:The pathogenesis of PAH is associated with multiple pathways such as the TGF-βsignaling pathway,PI3K-Akt signaling pathway,MAPK signaling pathway,HIF-1 signaling pathway and so on.The study of VEGFA,MAPK1,MAPK3,IL6,JUN and TNF are closely related to PAH is necessary for us to study further.Through gene interaction network and pathway analysis of disease-associated genes,which will help us to screen the critical target genes of PAH and provide a reference for clinical development of effective drugs for PAH.
文摘BACKGROUND Approximately half of all new cases of gastric cancer(GC)and related deaths occur in China.More than 80%of patients with GC are diagnosed at an advanced stage,which results in poor prognosis.Although HER2-directed therapy and immune checkpoint inhibitors have been somewhat successful,new drugs are still needed for the treatment of GC.Notably,several gene fusion-targeted drugs have been approved by the United States Food and Drug Administration for solid tumors,including GC,such as larotrectinib for NTRK fusion-positive cancers and zenocutuzumab for NRG1 fusion-positive cancers.However,gene fusions involving targetable genes have not been well characterized in Chinese patients with GC.AIM To identify the profile of fusions involving targetable genes in Chinese patients with GC using clinical specimens and determine the distribution of patients with gene fusion variants among the molecular subtypes of GC.METHODS We retrospectively analyzed gene fusion events in tumor tissue samples from 954 Chinese patients with GC.Clinicopathological characteristics were obtained from their medical records.Genetic alterations,such as single nucleotide variants,indels,amplifications,and gene fusions,were identified using a targeted sequencing panel containing 825 genes.Fusions were validated by fluorescence in situ hybridization(FISH)using break-apart probes.The microsatellite instability(MSI)status was evaluated using MSIsensor from the targeted sequencing panel data.Tumor mutational burden(TMB)was calculated using the total number of nonsynonymous mutations divided by the total genomic targeted region.Chi-square analysis was used to determine the enrichment of gene fusions associated with the molecular subtypes of GC.RESULTS We found that 1.68%(16/954)of patients harbored 20 fusion events involving targetable genes.RARA fusions(n=5)were the most common,followed by FGFR2,BRAF,MET,FGFR3,RET,ALK,EGFR,NTRK2,and NRG1 fusions.Two of the RARA fusions,EML4-ALK(E6:E20)and EGFRSEPTIN14(E7:E10),have been identified in other tumors but not in GC.Surprisingly,18 gene fusion events were previously not reported in any cancer types.Twelve of the eighteen novel gene fusions included complete exons encoding functional domains of targetable genes,such as the tyrosine kinase domain of receptor tyrosine kinases and the DNA-and ligand-binding domains of RARA.Consistent with the results of detection using the targeted sequencing fusion panel,the results of FISH(fluorescence in situ hybridization)confirmed the rearrangement of FGFR2 and BRAF in tumors from patients 04 and 09,respectively.Genetic analysis indicated that the fusion genes were significantly enriched in patients with ERBB2 amplification(P=0.02);however,there were no significant differences between fusion-positive and fusion-negative patients in age,sex,MSI status,and TMB.CONCLUSION We characterized the landscape of fusions involving targetable genes in a Chinese GC cohort and found that 1.68%of patients with GC harbor potential targetable gene fusions,which were enriched in patients with ERBB2 amplification.Gene fusion detection may provide a potential treatment strategy for patients with GC with disease progression following standard therapy.
基金supported by the National Key Research and Development Plan of China(2017YFD0501602)the Support Project of High-level Teachers in Beijing Municipal Universities in the Period of 13th Five Plan(IDHT20170516)
文摘The classic method for gene knockout (KO) is based on homologous recombination (HR) and embryonic stem cell technique (Gerlai,1996).Actually,the procedure of homologous replacement is complicated and time consuming,although it has been popular during the past decades.Recent years,genome editing which can cause DNA sequence-specific mutations in the genomes of cellular
基金National Nature Science Foundation of China(No.82060456)Hainan Provincial Key Research and Development Program Project Fund 405(No.ZDYF2019129)Hainan Provincial Postgraduate Innovation Research Project Fund(No.Hys2019-312)。
文摘Objective:To investigate the core target genes of miR-29b-3p,and analyze the clinical significance of the core target genes in glioma.Methods:Bioinformatics analysis was used to predict and screen the target genes of miR-29b-3p.STRING and Cytoscape software were used to analyze the protein-protein interaction(PPI)of target genes.the differences expression and survival prognosis in glioma were analyzed by GEPIA and CGGA.Independent prognostic factors analyzed by univariate and multivariate Cox proportional hazards regression model.Results:22 target genes of miR-29b-3p were predicted using LinkedOmics,miRDB,miRTarBase,TargetScan,and starbase databases.Through the construction of the PPI network,genes out of the network were removed,and a total of 16 genes were screened for further study of their clinical significance.Based on analysis of GEPIA and CGGA databases,COL2A1,DNMT3A,and DNMT3B were excluded.Through further analysis of the univariate and multivariate Cox proportional hazard regression model,finally identified three core target genes:SERPINH1,LOXL2,CDK6.Conclusion:Bioinformatics analysis showed that miR-29b-3p targeted three core genes such as SERPINH1,LOXL2,and CDK6 in glioma.The expression of these genes was different between brain normal tissues and gliomas,between different grades of tumor,IDH mutation status and 1p/19q codeletion status.Its high expression had adverse effects on overall survival and recurrence-free survival.These core target genes can be used as an independent prognostic factor.
基金supported by the grants of the Major State Basic Research Development Program (2006CB504403)the Supporting Program of the"Eleventh Five-year Plan"for Sci & Tech Research of China (2006BAD06A01 and 2006BAD06A12 )+1 种基金the Foundation Project of State key Laboratory of Veterinary Etiological Biology (SKLVEB2008ZZKT009)the Open Foundation Project of Guangdong Common Lab of Veterinary Public Health (GSKJ090202)
文摘Haemophilus parasuis is one kind of constant bacteria in porcine upper respiratory tract, and it can cause multiple serositis, arthritis and other diseases under certain conditions. Due to lack of efficient genetic operating system, its pathogenic mechanism is not very clear. Ligation with DNA ligase and fusion PCR were used to construct targeting hhdA gene of Haemophilus parasuis, respectively. The fidelity, application scope, operation and conditions of the constructed fusion fragments were compared. The results showed that construction with DNA ligase was more mature technology as manifested by more stable conditions and more extensive application. The fusion PCR method had high fidelity and simple operation, and the transformation rate was 9.5 times as high as that of ligation with DNA ligase. For this reason, this method was more suitable for construction of multi-fragment targeting genes. The study lays a foundation for establishing an efficient operating system of targeting gene of Haemophilus parasuis in the future.
基金Supported by National Natural Science Foundation of China(31260061,31060039)Key Laboratory of Special Biological Resource Development and Utilization of Universities in Yunnan Province(GXZD201601)+1 种基金Key Discipline Construction Project of Kunming UniversityNational College Students'Innovation Project of China
文摘[ Objective] This study aimed to screen target genes regulated by heat shock factor AtHsfAla in Arabidopsis thaliana. [ Method] Using AtHsfAla-in- serted mutant athsfala （SALK-068042） and wild-type A. thaliana seedlings as experimental materials, target genes regulated by heat shock factor AtHsfAla were screened by microarray assay. Differentially expressed genes were screened by multiple method. Specific functions of differentially expressed genes were analyzed by gene ontology （GO） analysis. Signal transduction pathways, in which differentia｜ly expressed genes were involved, were analyzed by pathway analysis. Gene-gene interaction network was constructed by Signal-Net. [ Result] A total of 3 672 differentially expressed genes were screened out. Up-regulated differentially expressed genes were involved in 198 functions and 7 signal transduction pathways; down-regulated differentially expressed genes were involved in 94 functions and 10 signal transduction pathways. In the signal transduction network, it was found that cwlNV4 and HXK3 had relatively high ability of mediation; AT1 G14240 and cwlNV4 ex- hibited the most interactions with other genes, which were located in key positions throughout the gene-gene interaction network. [ Conclusion] Heat shock factor AtHsfAla regulates a large number of target genes in A. thaliana.
基金Supported by Shandong Swine Industry Technology System and Science and Technology Planning Program for Basic Research in Qingdao City(12-1-4-14-jch)
文摘Gene targeting technology is an important means to investigate gene functions, but its efficiency of gene targeting is very low, especially for somatic cell targeting. Artificially induced double-strand breaks （DSB） and triplex forming oligonucleotide （TFO） are currently developed methods to improve the targeting efficiency. This paper summarized the basic principles, design ideas and application in gene targeting efficiency improvement of these two methods, analyzed and com- pared their characteristics, and finally proposed prospects for their future development.
