Clustered regularly interspaced short palindromic repeat sequences(CRISPR)and their accompanying proteins(Cas),commonly presenting in bacteria and archaea,make up the CRISPR/Cas system.As one of the funda-mental sourc...Clustered regularly interspaced short palindromic repeat sequences(CRISPR)and their accompanying proteins(Cas),commonly presenting in bacteria and archaea,make up the CRISPR/Cas system.As one of the funda-mental sources of nutrition for humans,edible crops play a crucial role in ensuring global food security.CRISPR/Cas9 gene editing has been applied to improve many crop traits,such as increasing nitrogen utilization efficiency,creating male sterile germplasm,and regulating tiller and spikelet formation.This paper provides a comprehensive overview of the use of CRISPR/Cas gene editing technology in crop genomes,covering the targeted genes,the types of editing that take place,the mechanism of action.Finally,we also discussed the efficiency of gene editing and pointed the future direction on how to speed up crop molecular breeding,increase breeding effectiveness,and produce more new crop varieties with high qualities.展开更多
The CRISPR/Cas technology is emerging as a revolutionary genome editing tool in diverse organisms including plants,and has quickly evolved into a suite of versatile tools for sequence-specific gene manipulations beyon...The CRISPR/Cas technology is emerging as a revolutionary genome editing tool in diverse organisms including plants,and has quickly evolved into a suite of versatile tools for sequence-specific gene manipulations beyond genome editing.Here,we review the most recent applications of the CRISPR/Cas toolkit in plants and also discuss key factors for improving CRISPR/Cas performance and strategies for reducing the off-target effects.Novel technical breakthroughs in mammalian research regarding the CRISPR/Cas toolkit will also be incorporated into this review in hope to stimulate prospective users from the plant research community to fully explore the potential of these technologies.展开更多
Based on rpoB gene micro array as target gene, we are going to use gene chip technology to test 24 mycobacterium standard specimens, 8 non-mycobacterium specimens and 86 mycobacterium clinical isolated specimens. As a...Based on rpoB gene micro array as target gene, we are going to use gene chip technology to test 24 mycobacterium standard specimens, 8 non-mycobacterium specimens and 86 mycobacterium clinical isolated specimens. As a result, after mycobacterium and non-mycobacterium standard specimens were duplicated by PCR, mycobacterium standard specimens reproduced 360bp DNA fragments; on the other hand, non-mycobacterium specimens did not reproduce any fragments except for hemolytic streptococcus and corynebacterium pseudodiphtheriticum which had the same results as mycobacterium standard specimens. Sensitive test is able to detect lpg tuberculosis mycobacterium DNA. The probe test showed that, among 21 oligonucleotide probes, probe-M. fortuitum and M. marinum were cross-hybrid; the other probes were specific. We used the new method to identify 126 mycobacterium clinical isolated specimens. The test results of this new method matched with conventional method. In conclusion, compared to the traditional method, the use of rpob gene chip technology to identify mycobacterium species will be faster, more accurate and higher value in application.展开更多
The headquarters of Plutus Financial Group Ltd,based in Hong Kong of China,stands as a silent yet razorsharp marker—rooted deeply in the heart of traditional finance,while piercing boundaries,exploring the vast nebul...The headquarters of Plutus Financial Group Ltd,based in Hong Kong of China,stands as a silent yet razorsharp marker—rooted deeply in the heart of traditional finance,while piercing boundaries,exploring the vast nebulae of blockchain and artificial intelligence.This integrated financial services group,newly listed on Nasdaq this February,is moving through the cut-and-thrust of the capital market with the postureof a"white knight."展开更多
AIM:To investigate the effect of in vivo environment on gene expression in Helicobacter pylori(H.pylori) as it relates to its survival in the host.METHODS:In vivo expression technology(IVET) systems are used to identi...AIM:To investigate the effect of in vivo environment on gene expression in Helicobacter pylori(H.pylori) as it relates to its survival in the host.METHODS:In vivo expression technology(IVET) systems are used to identify microbial virulence genes.We modified the IVET-transcriptional fusion vector,pIVET8,which uses antibiotic resistance as the basis for selection of candidate genes in host tissues to develop two unique IVET-promoter-screening vectors,pIVET11 and pIVET12.Our novel IVET systems were developed by the fusion of random Sau3A DNA fragments of H.pylori and a tandem-reporter system of chloramphenicol acetyltransferase and beta-galactosidase.Additionally,each vector contains a kanamycin resistance gene.We used a mouse macrophage cell line,RAW 264.7 and mice,as selective media to identify specific genes that H.pylori expresses in vivo.Gene expression studies were conducted by infecting RAW 264.7 cells with H.pylori.This was followed by real time polymerase chain reaction(PCR) analysis to determine the relative expression levels of in vivo induced genes.RESULTS:In this study,we have identified 31 in vivo induced(ivi) genes in the initial screens.These 31 genes belong to several functional gene families,including several well-known virulence factors that are expressed by the bacterium in infected mouse stomachs.Virulence factors,vacA and cagA,were found in this screen and are known to play important roles in H.pylori infection,colonization and pathogenesis.Their detection validates the efficacy of these screening systems.Some of the identified ivi genes have already been implicated to play an important role in the pathogenesis of H.pylori and other bacterial pathogens such as Escherichia coli and Vibrio cholerae.Transcription profiles of allivi genes were confirmed by real time PCR analysis of H.pylori RNA isolated from H.pylori infected RAW 264.7 macrophages.We compared the expression profile of H.pylori and RAW 264.7 coculture with that of H.pylori only.Some genes such as cag A,vac A,lpx C,mur I,tlp C,trx B,sod B,tnp B,pgi,rbf A and inf B showed a 2-20 fold upregulation.Statistically significant upregulation was obtained for all the above mentioned genes(P < 0.05).tlp C,cag A,vac A,sod B,rbf A,inf B,tnp B,lpx C and mur I were also significantly upregulated(P < 0.01).These data suggest a strong correlation between results obtained in vitro in the macrophage cell line and in the intact animal.CONCLUSION:The positive identification of these genes demonstrates that our IVET systems are powerful tools for studying H.pylori gene expression in the host environment.展开更多
The differential expression of genes in HepG2 cells caused by UC001 kfo RNAi was investigated using RNA-seq. HepG2 cells were infected by Lenti-sh UC001 kfo lentivirus particles. The expression of UC001 kfo m RNA in t...The differential expression of genes in HepG2 cells caused by UC001 kfo RNAi was investigated using RNA-seq. HepG2 cells were infected by Lenti-sh UC001 kfo lentivirus particles. The expression of UC001 kfo m RNA in the HepG2-sh UC001 kfo cell line was detected by real-time PCR. RNA-seq technology was used to identify the difference in the expression of genes regulated by lnc RNA UC001 kfo in the HepG2 cell line. Gene ontology and signaling pathway analysis were performed to reveal the biological functions of the genes encoding of significantly different m RNAs. The results showed that m RNAs were differentially expressed between the HepG2-sh UC001 kfo cell line and the HepG2 cell line. The UC001 kfo m RNA was significantly down-regulated in the stable cell line HepG2-sh UC001kfo（P〈0.001）. Additionally, we found 19 signaling pathways or functional classifications encompassing 30 genes that played a role in cancer characteristics, cell adhesion, invasion and migration. The results also showed that the expression of many genes associated with cancer cell invasion and metastasis was decreased with the down-regulation of the lnc RNA UC001 kfo. Lnc RNA UC001 kfo may play a role in regulating cancer cell invasion and metastasis. It was suggested that m RNAs were differentially expressed in the HepG2 cell line after the down-regulation of lnc RNA-UC001 kfo. Some took part in the extracellular matrix, cell adhesion, motility, growth, and localization. The genes encoding of differentially expressed m RNAs may participate in cell invasion and metastasis.展开更多
Gene expression must be precisely regulated in cells and functional nucleic acids are the most widely utilized tools for gene manipulation.Photocontrol of how these nucleic acid tools work in the cellular environment ...Gene expression must be precisely regulated in cells and functional nucleic acids are the most widely utilized tools for gene manipulation.Photocontrol of how these nucleic acid tools work in the cellular environment can precisely manipulate gene expression through a non-invasive way.Here we report a methodology on multiplex photocontrol of functional nucleic acids to achieve totally temporal and orthogonal regulation of gene expression in living cells.We select two functional nucleic acid systems as examples,DNAzyme and CRISPR/Cas9,and demonstrate the power of light control for precise gene manipulation by rational design of chemically modified oligonucleotides through introduction of two photocleavable linkers.Unlike the previous modulation of functional nucleic acids by simply activating or deactivating,we successfully achieve versatile controlling patterns using light as the governing factor.This design represents a generalized pathway towards the photo-controllable functional nucleic acids,which greatly enriches the toolbox for optogenetic studies.展开更多
This study investigated the effects of the luxS gene on the biofilm formation of Lactobacillus fermentum and its stress resistance.Deletion of the luxS gene was related to the physiological characteristics of biofilm ...This study investigated the effects of the luxS gene on the biofilm formation of Lactobacillus fermentum and its stress resistance.Deletion of the luxS gene was related to the physiological characteristics of biofilm development,strain resistance,bacterial morphology,biofilm morphology,functional group types,the main components of the biofilm,and the expression of related genes.Therefore,a luxS gene-deficient strain was constructed usingλRed gene recombination technology,and the biofilm production and stress resistance of the WT andΔluxS strains were compared.Deletion of the gene reduced the biofilm formation of L.fermentum and its resistance to acid,bile salt,high temperatures,and a hypertonic environment.Further,this work found that its deletion affects the types of functional groups in biofilms,and at the same time downregulate the expressions of the fabI,cysE,argR,and purD genes,thus reducing the levels of fatty acids,exopolysaccharide,protein,and eDNA in biofilms.A correlation analysis found that protein in biofilms was the most important factor affecting biofilm formation.In conclusion,the luxS gene in L.fermentum is involved in the regulation of biofilm formation and stress resistance.These findings provide a theoretical basis for the study of the mechanism of biofilm formation in beneficial bacteria.展开更多
BACKGROUND The aim of this study was to investigate the complex heterozygous mutations of ANK1 and SPTA1 in the same individual and improve our understanding of hereditary spherocytosis(HS)in children.We also hope to ...BACKGROUND The aim of this study was to investigate the complex heterozygous mutations of ANK1 and SPTA1 in the same individual and improve our understanding of hereditary spherocytosis(HS)in children.We also hope to promote the application of gene detection technology in children with HS,with the goals of identifying more related gene mutations,supporting the acquisition of improved molecular genetic information to further reveal the pathogenesis of HS in children,and providing important guidance for the diagnosis,treatment,and prevention of HS in children.CASE SUMMARY A 1-year and 5-month-old patient presented jaundice during the neonatal period,mild anemia 8 months later,splenic enlargement at 1 year and 5 months,and brittle red blood cell permeability.Genetic testing was performed on the patient,their parents,and sister.Swiss Model software was used to predict the protein structure of complex heterozygous mutations in ANK1 and SPTA1.Genetic testing revealed that the patient harbored a new mutation in the ANK1 gene from the father and a mutation in the SPTA1 gene from the mother.Combined with the clinical symptoms of the children,it is suggested that the newly discovered complex heterozygous mutations of ANK1 and SPTA1 may be the cause,providing important guidance for revealing the pathogenesis,diagnosis,treatment,and promotion of gene detection technology in children with HS.CONCLUSION This case involves an unreported complex heterozygous mutation of ANK1 and SPTA1,which provides a reference for exploring HS.展开更多
This paper reviews the latest advances and applications of gene editing technologies in the study of aging-related genes.In recent years,gene editing has achieved significant progress in the biomedical field,with cont...This paper reviews the latest advances and applications of gene editing technologies in the study of aging-related genes.In recent years,gene editing has achieved significant progress in the biomedical field,with continual improvements in accuracy and efficiency.Gene editing technologies demonstrate unique advantages in aging research,providing powerful tools for elucidating aging mechanisms and developing anti-aging interventions.This paper offers a detailed overview of major gene editing technologies(such as the CRISPR/Cas system,TALENs,and ZFNs)as well as emerging editing techniques(including base editing,prime editing,and epigenetic editing),describing their principles and applications.It also discusses research progress in areas such as constructing aging gene models,disease models,in vivo editing,and in vitro editing.Furthermore,it analyzes current technological challenges and proposes corresponding optimization strategies.Finally,it considers future directions for the development of gene editing technologies in aging research,including technological innovation and integrated multi-technology applications.Through these advances and innovations,gene editing is expected to play an increasingly important role in the anti-aging field,offering new strategies and methods to promote human health and longevity.展开更多
Environmental pollution and the spread of pathogenic microorganisms pose a significant threat to the health of humans and the planet.Thus,understanding and detecting microorganisms is crucial for maintaining a healthy...Environmental pollution and the spread of pathogenic microorganisms pose a significant threat to the health of humans and the planet.Thus,understanding and detecting microorganisms is crucial for maintaining a healthy living environment.Nanopore sequencing is a single-molecule detection method developed in the 1990s that has revolutionized various research fields.It offers several advantages over traditional sequencing methods,including low cost,label-free,time-saving detection speed,long sequencing reading,real-time monitoring,convenient carrying,and other significant advantages.In this review,we summarize the technical principles and characteristics of nanopore sequencing and discuss its applications in amplicon sequencing,metagenome sequencing,and whole-genome sequencing of environmental microorganisms,as well as its in situ application under some special circumstances.We also analyze the advantages and challenges of nanopore sequencing in microbiology research.Overall,nanopore sequencing has the potential to greatly enhance the detection and understanding of microorganisms in environmental research,but further developments are needed to overcome the current challenges.展开更多
Background Gastric cancer ranks high among the most common causes of cancer-related death worldwide. This study was designed to explore key genes involved in the progression of normal gastric epithelial cells to moder...Background Gastric cancer ranks high among the most common causes of cancer-related death worldwide. This study was designed to explore key genes involved in the progression of normal gastric epithelial cells to moderate gastric epithelial dysplasia （mGED） and to gastric cancer. Methods Twelve pairs of mGED tissues, gastric cancer tissues, and normal gastric tissues were collected by gastroscopy. Total RNA was then extracted and purified. After the addition of fluorescent tags, hybridization was carried out on a Gene chip microarray slide. Significance analysis of microarrays was performed to determine significant differences in gene expression between the different tissue types. Results Microarray data analysis revealed totally 34 genes that were expressed differently： 18 highly expressed （fold change 〉2; P 〈0.01） and 16 down-regulated （fold change 〉2; P 〈0.01）. Of the 34 genes, 24 belonged to several different functional categories such as structural molecule activity, extracellular regions, structural formation, cell death, biological adhesion, developmental processes, locomotion, and biological regulation that were associated with cancer. The remaining 10 genes were not involved in cancer research. Of these genes, the expression levels of Matrix metalloproteinase-12 （MMP12）, Caspase-associated recruitment domain 14 （CARD14）, and Chitinase 3-like 1 （CHI3L1） were confirmed by semi-quantitative RT-PCR. A two-way clustering algorithm divided the 36 samples into three categories and the overall correct classification efficiency was 80.6% （29/36）. Almost all of these genes （31/34） showed constant changes in the process of normal gastric epithelial cells to mGED to gastric cancer. Conclusions The results of this study provided global gene expression profiles during the development and progression from normal gastric epithelial cells to mGED to gastric cancer. These data may provide new insights into the molecular pathology of gastric cancer which may be useful for the detection, diagnosis, and treatment.展开更多
Comprehensive Summary Currently,CRISPR/Cas9 technology has found widespread applications across various domains.However,the utility of CRISPR/Cas9 is encumbered by issues pertaining to its reliability and safety,prima...Comprehensive Summary Currently,CRISPR/Cas9 technology has found widespread applications across various domains.However,the utility of CRISPR/Cas9 is encumbered by issues pertaining to its reliability and safety,primarily stemming from the uncontrolled activity of the system.Therefore,the design and development of CRISPR/Cas9 systems with controllable activity is of paramount importance.Biotin,characterized by its small molecular weight,and streptavidin,distinguished by its substantial spatial steric hindrance,can be harnessed as an ideal OFF switch(termed a"bioactivity brake")due to their interaction characteristics.In this work,we present a strategy that employs the streptavidin-biotin interaction as a"brake system"for CRISPR/Cas9,effectively allowing for the shutdown of the enzymatic activity of CRISPR/Cas9.展开更多
The CRISPR/Cas(clustered regularly interspaced short palindromic repeats/CRISPRassociated proteins) system was first identified in bacteria and archaea and can degrade exogenous substrates. It was developed as a gene ...The CRISPR/Cas(clustered regularly interspaced short palindromic repeats/CRISPRassociated proteins) system was first identified in bacteria and archaea and can degrade exogenous substrates. It was developed as a gene editing technology in 2013. Over the subsequent years, it has received extensive attention owing to its easy manipulation, high efficiency, and wide application in gene mutation and transcriptional regulation in mammals and plants. The process of CRISPR/Cas is optimized constantly and its application has also expanded dramatically. Therefore, CRISPR/Cas is considered a revolutionary technology in plant biology. Here, we introduce the mechanism of the type II CRISPR/Cas called CRISPR/Cas9, update its recent advances in various applications in plants, and discuss its future prospects to provide an argument for its use in the study of medicinal plants.展开更多
基金supported by Jilin Provincial Department of Education(JKH20230394KJ).
文摘Clustered regularly interspaced short palindromic repeat sequences(CRISPR)and their accompanying proteins(Cas),commonly presenting in bacteria and archaea,make up the CRISPR/Cas system.As one of the funda-mental sources of nutrition for humans,edible crops play a crucial role in ensuring global food security.CRISPR/Cas9 gene editing has been applied to improve many crop traits,such as increasing nitrogen utilization efficiency,creating male sterile germplasm,and regulating tiller and spikelet formation.This paper provides a comprehensive overview of the use of CRISPR/Cas gene editing technology in crop genomes,covering the targeted genes,the types of editing that take place,the mechanism of action.Finally,we also discussed the efficiency of gene editing and pointed the future direction on how to speed up crop molecular breeding,increase breeding effectiveness,and produce more new crop varieties with high qualities.
基金supported by the National Natural Science Foundation of China(Grant Nos. 31522006 and 31570276)funds from Sun Yatsen University and from China's Thousand Young Talents Program to J.-F.Li
文摘The CRISPR/Cas technology is emerging as a revolutionary genome editing tool in diverse organisms including plants,and has quickly evolved into a suite of versatile tools for sequence-specific gene manipulations beyond genome editing.Here,we review the most recent applications of the CRISPR/Cas toolkit in plants and also discuss key factors for improving CRISPR/Cas performance and strategies for reducing the off-target effects.Novel technical breakthroughs in mammalian research regarding the CRISPR/Cas toolkit will also be incorporated into this review in hope to stimulate prospective users from the plant research community to fully explore the potential of these technologies.
文摘Based on rpoB gene micro array as target gene, we are going to use gene chip technology to test 24 mycobacterium standard specimens, 8 non-mycobacterium specimens and 86 mycobacterium clinical isolated specimens. As a result, after mycobacterium and non-mycobacterium standard specimens were duplicated by PCR, mycobacterium standard specimens reproduced 360bp DNA fragments; on the other hand, non-mycobacterium specimens did not reproduce any fragments except for hemolytic streptococcus and corynebacterium pseudodiphtheriticum which had the same results as mycobacterium standard specimens. Sensitive test is able to detect lpg tuberculosis mycobacterium DNA. The probe test showed that, among 21 oligonucleotide probes, probe-M. fortuitum and M. marinum were cross-hybrid; the other probes were specific. We used the new method to identify 126 mycobacterium clinical isolated specimens. The test results of this new method matched with conventional method. In conclusion, compared to the traditional method, the use of rpob gene chip technology to identify mycobacterium species will be faster, more accurate and higher value in application.
文摘The headquarters of Plutus Financial Group Ltd,based in Hong Kong of China,stands as a silent yet razorsharp marker—rooted deeply in the heart of traditional finance,while piercing boundaries,exploring the vast nebulae of blockchain and artificial intelligence.This integrated financial services group,newly listed on Nasdaq this February,is moving through the cut-and-thrust of the capital market with the postureof a"white knight."
基金Supported by Intramural Research Program of the National Institutes of Health,National Institute of Diabetes and Digestive and Kidney DiseaseThe Division of Intramural Research of the National Institute of Allergy and Infectious DiseasesAn Inter-Agency Agreement (Y3-DK-3521-07) with the National Institute on Minority Health and Health Disparities
文摘AIM:To investigate the effect of in vivo environment on gene expression in Helicobacter pylori(H.pylori) as it relates to its survival in the host.METHODS:In vivo expression technology(IVET) systems are used to identify microbial virulence genes.We modified the IVET-transcriptional fusion vector,pIVET8,which uses antibiotic resistance as the basis for selection of candidate genes in host tissues to develop two unique IVET-promoter-screening vectors,pIVET11 and pIVET12.Our novel IVET systems were developed by the fusion of random Sau3A DNA fragments of H.pylori and a tandem-reporter system of chloramphenicol acetyltransferase and beta-galactosidase.Additionally,each vector contains a kanamycin resistance gene.We used a mouse macrophage cell line,RAW 264.7 and mice,as selective media to identify specific genes that H.pylori expresses in vivo.Gene expression studies were conducted by infecting RAW 264.7 cells with H.pylori.This was followed by real time polymerase chain reaction(PCR) analysis to determine the relative expression levels of in vivo induced genes.RESULTS:In this study,we have identified 31 in vivo induced(ivi) genes in the initial screens.These 31 genes belong to several functional gene families,including several well-known virulence factors that are expressed by the bacterium in infected mouse stomachs.Virulence factors,vacA and cagA,were found in this screen and are known to play important roles in H.pylori infection,colonization and pathogenesis.Their detection validates the efficacy of these screening systems.Some of the identified ivi genes have already been implicated to play an important role in the pathogenesis of H.pylori and other bacterial pathogens such as Escherichia coli and Vibrio cholerae.Transcription profiles of allivi genes were confirmed by real time PCR analysis of H.pylori RNA isolated from H.pylori infected RAW 264.7 macrophages.We compared the expression profile of H.pylori and RAW 264.7 coculture with that of H.pylori only.Some genes such as cag A,vac A,lpx C,mur I,tlp C,trx B,sod B,tnp B,pgi,rbf A and inf B showed a 2-20 fold upregulation.Statistically significant upregulation was obtained for all the above mentioned genes(P < 0.05).tlp C,cag A,vac A,sod B,rbf A,inf B,tnp B,lpx C and mur I were also significantly upregulated(P < 0.01).These data suggest a strong correlation between results obtained in vitro in the macrophage cell line and in the intact animal.CONCLUSION:The positive identification of these genes demonstrates that our IVET systems are powerful tools for studying H.pylori gene expression in the host environment.
基金supported by National Natural Science Foudation of China(No.U1404309)
文摘The differential expression of genes in HepG2 cells caused by UC001 kfo RNAi was investigated using RNA-seq. HepG2 cells were infected by Lenti-sh UC001 kfo lentivirus particles. The expression of UC001 kfo m RNA in the HepG2-sh UC001 kfo cell line was detected by real-time PCR. RNA-seq technology was used to identify the difference in the expression of genes regulated by lnc RNA UC001 kfo in the HepG2 cell line. Gene ontology and signaling pathway analysis were performed to reveal the biological functions of the genes encoding of significantly different m RNAs. The results showed that m RNAs were differentially expressed between the HepG2-sh UC001 kfo cell line and the HepG2 cell line. The UC001 kfo m RNA was significantly down-regulated in the stable cell line HepG2-sh UC001kfo（P〈0.001）. Additionally, we found 19 signaling pathways or functional classifications encompassing 30 genes that played a role in cancer characteristics, cell adhesion, invasion and migration. The results also showed that the expression of many genes associated with cancer cell invasion and metastasis was decreased with the down-regulation of the lnc RNA UC001 kfo. Lnc RNA UC001 kfo may play a role in regulating cancer cell invasion and metastasis. It was suggested that m RNAs were differentially expressed in the HepG2 cell line after the down-regulation of lnc RNA-UC001 kfo. Some took part in the extracellular matrix, cell adhesion, motility, growth, and localization. The genes encoding of differentially expressed m RNAs may participate in cell invasion and metastasis.
基金supported by the National Natural Science Foundation of China(22222706,22307107,22477144)the National Key R&D Program of China(2020YFA0211200)the Guangdong Basic Research Center of Excellence(GBRCE)for Functional Molecular Engineering and the Guangdong Basic and Applied Basic Research Foundation(2024B1515040028)。
文摘Gene expression must be precisely regulated in cells and functional nucleic acids are the most widely utilized tools for gene manipulation.Photocontrol of how these nucleic acid tools work in the cellular environment can precisely manipulate gene expression through a non-invasive way.Here we report a methodology on multiplex photocontrol of functional nucleic acids to achieve totally temporal and orthogonal regulation of gene expression in living cells.We select two functional nucleic acid systems as examples,DNAzyme and CRISPR/Cas9,and demonstrate the power of light control for precise gene manipulation by rational design of chemically modified oligonucleotides through introduction of two photocleavable linkers.Unlike the previous modulation of functional nucleic acids by simply activating or deactivating,we successfully achieve versatile controlling patterns using light as the governing factor.This design represents a generalized pathway towards the photo-controllable functional nucleic acids,which greatly enriches the toolbox for optogenetic studies.
基金supported by National Natural Science Foundation of China(No.31960467)the Programs of Natural Science Foundation of Inner Mongolia(No.2019BS03002)the Research and Innovation Projects for Postgraduates of Inner Mongolia(B20191147Z).
文摘This study investigated the effects of the luxS gene on the biofilm formation of Lactobacillus fermentum and its stress resistance.Deletion of the luxS gene was related to the physiological characteristics of biofilm development,strain resistance,bacterial morphology,biofilm morphology,functional group types,the main components of the biofilm,and the expression of related genes.Therefore,a luxS gene-deficient strain was constructed usingλRed gene recombination technology,and the biofilm production and stress resistance of the WT andΔluxS strains were compared.Deletion of the gene reduced the biofilm formation of L.fermentum and its resistance to acid,bile salt,high temperatures,and a hypertonic environment.Further,this work found that its deletion affects the types of functional groups in biofilms,and at the same time downregulate the expressions of the fabI,cysE,argR,and purD genes,thus reducing the levels of fatty acids,exopolysaccharide,protein,and eDNA in biofilms.A correlation analysis found that protein in biofilms was the most important factor affecting biofilm formation.In conclusion,the luxS gene in L.fermentum is involved in the regulation of biofilm formation and stress resistance.These findings provide a theoretical basis for the study of the mechanism of biofilm formation in beneficial bacteria.
基金Supported by The Science and Technology Department of Sichuan Province,No.2021JDKP0015.
文摘BACKGROUND The aim of this study was to investigate the complex heterozygous mutations of ANK1 and SPTA1 in the same individual and improve our understanding of hereditary spherocytosis(HS)in children.We also hope to promote the application of gene detection technology in children with HS,with the goals of identifying more related gene mutations,supporting the acquisition of improved molecular genetic information to further reveal the pathogenesis of HS in children,and providing important guidance for the diagnosis,treatment,and prevention of HS in children.CASE SUMMARY A 1-year and 5-month-old patient presented jaundice during the neonatal period,mild anemia 8 months later,splenic enlargement at 1 year and 5 months,and brittle red blood cell permeability.Genetic testing was performed on the patient,their parents,and sister.Swiss Model software was used to predict the protein structure of complex heterozygous mutations in ANK1 and SPTA1.Genetic testing revealed that the patient harbored a new mutation in the ANK1 gene from the father and a mutation in the SPTA1 gene from the mother.Combined with the clinical symptoms of the children,it is suggested that the newly discovered complex heterozygous mutations of ANK1 and SPTA1 may be the cause,providing important guidance for revealing the pathogenesis,diagnosis,treatment,and promotion of gene detection technology in children with HS.CONCLUSION This case involves an unreported complex heterozygous mutation of ANK1 and SPTA1,which provides a reference for exploring HS.
文摘This paper reviews the latest advances and applications of gene editing technologies in the study of aging-related genes.In recent years,gene editing has achieved significant progress in the biomedical field,with continual improvements in accuracy and efficiency.Gene editing technologies demonstrate unique advantages in aging research,providing powerful tools for elucidating aging mechanisms and developing anti-aging interventions.This paper offers a detailed overview of major gene editing technologies(such as the CRISPR/Cas system,TALENs,and ZFNs)as well as emerging editing techniques(including base editing,prime editing,and epigenetic editing),describing their principles and applications.It also discusses research progress in areas such as constructing aging gene models,disease models,in vivo editing,and in vitro editing.Furthermore,it analyzes current technological challenges and proposes corresponding optimization strategies.Finally,it considers future directions for the development of gene editing technologies in aging research,including technological innovation and integrated multi-technology applications.Through these advances and innovations,gene editing is expected to play an increasingly important role in the anti-aging field,offering new strategies and methods to promote human health and longevity.
基金grateful to the financial support from the National Natural Science Foundation of China(Nos.22025407,21974144)Institute of Chemistry,Chinese Academy of Sciences。
文摘Environmental pollution and the spread of pathogenic microorganisms pose a significant threat to the health of humans and the planet.Thus,understanding and detecting microorganisms is crucial for maintaining a healthy living environment.Nanopore sequencing is a single-molecule detection method developed in the 1990s that has revolutionized various research fields.It offers several advantages over traditional sequencing methods,including low cost,label-free,time-saving detection speed,long sequencing reading,real-time monitoring,convenient carrying,and other significant advantages.In this review,we summarize the technical principles and characteristics of nanopore sequencing and discuss its applications in amplicon sequencing,metagenome sequencing,and whole-genome sequencing of environmental microorganisms,as well as its in situ application under some special circumstances.We also analyze the advantages and challenges of nanopore sequencing in microbiology research.Overall,nanopore sequencing has the potential to greatly enhance the detection and understanding of microorganisms in environmental research,but further developments are needed to overcome the current challenges.
文摘Background Gastric cancer ranks high among the most common causes of cancer-related death worldwide. This study was designed to explore key genes involved in the progression of normal gastric epithelial cells to moderate gastric epithelial dysplasia （mGED） and to gastric cancer. Methods Twelve pairs of mGED tissues, gastric cancer tissues, and normal gastric tissues were collected by gastroscopy. Total RNA was then extracted and purified. After the addition of fluorescent tags, hybridization was carried out on a Gene chip microarray slide. Significance analysis of microarrays was performed to determine significant differences in gene expression between the different tissue types. Results Microarray data analysis revealed totally 34 genes that were expressed differently： 18 highly expressed （fold change 〉2; P 〈0.01） and 16 down-regulated （fold change 〉2; P 〈0.01）. Of the 34 genes, 24 belonged to several different functional categories such as structural molecule activity, extracellular regions, structural formation, cell death, biological adhesion, developmental processes, locomotion, and biological regulation that were associated with cancer. The remaining 10 genes were not involved in cancer research. Of these genes, the expression levels of Matrix metalloproteinase-12 （MMP12）, Caspase-associated recruitment domain 14 （CARD14）, and Chitinase 3-like 1 （CHI3L1） were confirmed by semi-quantitative RT-PCR. A two-way clustering algorithm divided the 36 samples into three categories and the overall correct classification efficiency was 80.6% （29/36）. Almost all of these genes （31/34） showed constant changes in the process of normal gastric epithelial cells to mGED to gastric cancer. Conclusions The results of this study provided global gene expression profiles during the development and progression from normal gastric epithelial cells to mGED to gastric cancer. These data may provide new insights into the molecular pathology of gastric cancer which may be useful for the detection, diagnosis, and treatment.
基金the National Natural Science Foundation of China(Nos.22177089,21721005,92153303,22037004,22177088)the Fundamental Research Funds for the Central Universities(2042023kf0204)Translational Medicine and Interdisciplinary Research Joint Fund of Zhongnan Hospital of Wuhan University(Grant No.ZNJC202309).
文摘Comprehensive Summary Currently,CRISPR/Cas9 technology has found widespread applications across various domains.However,the utility of CRISPR/Cas9 is encumbered by issues pertaining to its reliability and safety,primarily stemming from the uncontrolled activity of the system.Therefore,the design and development of CRISPR/Cas9 systems with controllable activity is of paramount importance.Biotin,characterized by its small molecular weight,and streptavidin,distinguished by its substantial spatial steric hindrance,can be harnessed as an ideal OFF switch(termed a"bioactivity brake")due to their interaction characteristics.In this work,we present a strategy that employs the streptavidin-biotin interaction as a"brake system"for CRISPR/Cas9,effectively allowing for the shutdown of the enzymatic activity of CRISPR/Cas9.
文摘The CRISPR/Cas(clustered regularly interspaced short palindromic repeats/CRISPRassociated proteins) system was first identified in bacteria and archaea and can degrade exogenous substrates. It was developed as a gene editing technology in 2013. Over the subsequent years, it has received extensive attention owing to its easy manipulation, high efficiency, and wide application in gene mutation and transcriptional regulation in mammals and plants. The process of CRISPR/Cas is optimized constantly and its application has also expanded dramatically. Therefore, CRISPR/Cas is considered a revolutionary technology in plant biology. Here, we introduce the mechanism of the type II CRISPR/Cas called CRISPR/Cas9, update its recent advances in various applications in plants, and discuss its future prospects to provide an argument for its use in the study of medicinal plants.
