Reverse Genetic Approaches in Zebrafish  

作　　者：Peng Huang Zuoyan Zhu Shuo Lin Bo Zhang 

机构地区：[1]Key Laboratory of Cell Proliferation and Differentiation of Ministry of Education,College of Life Sciences,Peking University,Beijing 100871,China [2]Department of Molecular,Cell&Developmental Biology,University of California,Los Angeles,CA 90095,USA

出　　处：《Journal of Genetics and Genomics》2012年第9期421-433,共13页遗传学报（英文版）

基　　金：partially supported by the grants from National Program on Key Basic Research Project(973 program)(Nos.2012CB945101 and 201 ICBAO 1000);National Natural Science Foundation of China(NSFC)(Nos. 31110103904 and 30730056)

摘　　要：Zebrafish（Danio rerio） is a well-established vertebrate animal model.A comprehensive collection of reverse genetics tools has been developed for studying gene function in this useful organism.Morpholino is the most widely used reagent to knock down target gene expression post-transcriptionally.For a long time,targeted genome modification has been heavily relied on large-scale traditional forward genetic screens,such as ENU（N-ethyl-N-nitrosourea） mutagenesis derived TILLING（Targeting Induced Local Lesions IN Genomes） strategy and pseudo-typed retrovirus mediated insertional mutagenesis.Recently,engineered endonucleases,including ZFNs（zinc finger nucleases） and TALENs（transcription activator-like effector nucleases）,provide new and efficient strategies to directly generate site-specific indel mutations by inducing double strand breaks in target genes.Here we summarize the major reverse genetic approaches for loss-of-function studies used and emerging in zebrafish,including strategies based on genome-wide mutagenesis and methods for site-specific gene targeting.Future directions and expectations will also be discussed.Zebrafish（Danio rerio） is a well-established vertebrate animal model.A comprehensive collection of reverse genetics tools has been developed for studying gene function in this useful organism.Morpholino is the most widely used reagent to knock down target gene expression post-transcriptionally.For a long time,targeted genome modification has been heavily relied on large-scale traditional forward genetic screens,such as ENU（N-ethyl-N-nitrosourea） mutagenesis derived TILLING（Targeting Induced Local Lesions IN Genomes） strategy and pseudo-typed retrovirus mediated insertional mutagenesis.Recently,engineered endonucleases,including ZFNs（zinc finger nucleases） and TALENs（transcription activator-like effector nucleases）,provide new and efficient strategies to directly generate site-specific indel mutations by inducing double strand breaks in target genes.Here we summarize the major reverse genetic approaches for loss-of-function studies used and emerging in zebrafish,including strategies based on genome-wide mutagenesis and methods for site-specific gene targeting.Future directions and expectations will also be discussed.

关 键 词：Zebrafish Reverse genetics Morpholino TILLING Retrovirus ZFN TALEN Gene targeting 

分 类 号：Q943[生物学—植物学] X503.225[环境科学与工程—环境工程]