Deciphering gene function is fundamental to engineering of microbiology.The clustered regularly interspaced short palindromic repeats(CRISPR)system has been adapted for gene repression across a range of hosts,creating...Deciphering gene function is fundamental to engineering of microbiology.The clustered regularly interspaced short palindromic repeats(CRISPR)system has been adapted for gene repression across a range of hosts,creating a versatile tool called CRISPR interference(CRISPRi)that enables genome-scale analysis of gene function.This approach has yielded significant advances in the design of genome-scale CRISPRi libraries,as well as in applica-tions of CRISPRi screening in medical and industrial microbiology.This review provides an overview of the recent progress made in pooled and arrayed CRISPRi screening in microorganisms and highlights representative studies that have employed this method.Additionally,the challenges associated with CRISPRi screening are discussed,and potential solutions for optimizing this strategy are proposed.展开更多
G protein-coupled receptors(GPCRs)are pivotal in mediating diverse physiological and pathological processes,rendering them promising targets for drug discovery.GPCRs account for about 40%of FDA-approved drugs,represen...G protein-coupled receptors(GPCRs)are pivotal in mediating diverse physiological and pathological processes,rendering them promising targets for drug discovery.GPCRs account for about 40%of FDA-approved drugs,representing the most successful drug targets.However,only approximately 15%of the 800 human GPCRs are targeted by market drugs,leaving numerous opportunities for drug discovery among the remaining receptors.Cell expression systems play crucial roles in the GPCR drug discovery field,including novel target identification,structural and functional characterization,potential ligand screening,signal pathway elucidation,and drug safety evaluation.Here,we discuss the principles,applications,and limitations of widely used cell expression systems in GPCR-targeted drug discovery,GPCR function investigation,signal pathway characterization,and pharmacological property studies.We also propose three strategies for constructing genome-wide pan-GPCR cell libraries,which will provide a powerful platform for GPCR ligand screening,and facilitate the study of GPCR mechanisms and drug safety evaluation,ultimately accelerating the process of GPCR-targeted drug discovery.展开更多
功能性基因的筛选是探索生物进程、研究疾病发生发展和诠释基因功能的重要方法,在生物、医药、新治疗靶点筛选及肿瘤耐药等方面有广泛的应用。CRISPR-Cas9(Clustered regularly interspaced short palindromic repeat sequences/CRISPR-...功能性基因的筛选是探索生物进程、研究疾病发生发展和诠释基因功能的重要方法,在生物、医药、新治疗靶点筛选及肿瘤耐药等方面有广泛的应用。CRISPR-Cas9(Clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein 9)技术作为近期热门的基因编辑工具,能够高通量地对基因组进行精准修饰,为实现功能性基因的筛选提供了简便高效的技术支持。文中对CRISPR-cas9技术应用于功能性基因筛选的方法及研究进展进行了综述。展开更多
基金supported by the National Key R&D Program of China(2018YFA0901500)the National Natural Science Foundation of China(32222004 and 32270101)the Youth Innovation Promotion Association of Chinese Academy of Sciences(2021177).
文摘Deciphering gene function is fundamental to engineering of microbiology.The clustered regularly interspaced short palindromic repeats(CRISPR)system has been adapted for gene repression across a range of hosts,creating a versatile tool called CRISPR interference(CRISPRi)that enables genome-scale analysis of gene function.This approach has yielded significant advances in the design of genome-scale CRISPRi libraries,as well as in applica-tions of CRISPRi screening in medical and industrial microbiology.This review provides an overview of the recent progress made in pooled and arrayed CRISPRi screening in microorganisms and highlights representative studies that have employed this method.Additionally,the challenges associated with CRISPRi screening are discussed,and potential solutions for optimizing this strategy are proposed.
基金supported by introducing the talented person scientific research starts funds subsidization project of Chengdu University of Traditional Chinese Medicine(030040019,030040017,China).
文摘G protein-coupled receptors(GPCRs)are pivotal in mediating diverse physiological and pathological processes,rendering them promising targets for drug discovery.GPCRs account for about 40%of FDA-approved drugs,representing the most successful drug targets.However,only approximately 15%of the 800 human GPCRs are targeted by market drugs,leaving numerous opportunities for drug discovery among the remaining receptors.Cell expression systems play crucial roles in the GPCR drug discovery field,including novel target identification,structural and functional characterization,potential ligand screening,signal pathway elucidation,and drug safety evaluation.Here,we discuss the principles,applications,and limitations of widely used cell expression systems in GPCR-targeted drug discovery,GPCR function investigation,signal pathway characterization,and pharmacological property studies.We also propose three strategies for constructing genome-wide pan-GPCR cell libraries,which will provide a powerful platform for GPCR ligand screening,and facilitate the study of GPCR mechanisms and drug safety evaluation,ultimately accelerating the process of GPCR-targeted drug discovery.
文摘功能性基因的筛选是探索生物进程、研究疾病发生发展和诠释基因功能的重要方法,在生物、医药、新治疗靶点筛选及肿瘤耐药等方面有广泛的应用。CRISPR-Cas9(Clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein 9)技术作为近期热门的基因编辑工具,能够高通量地对基因组进行精准修饰,为实现功能性基因的筛选提供了简便高效的技术支持。文中对CRISPR-cas9技术应用于功能性基因筛选的方法及研究进展进行了综述。
