Current perspectives in zebrafish reverse genetics: moving forward.
Use of the zebrafish as a model of vertebrate development and disease has expanded dramatically over the past decade. While many articles have discussed the strengths of zebrafish forward genetics (the phenotype-driven approach), there has been less emphasis on equally important and frequently used reverse genetics (the candidate gene-driven approach). Here we review both current and prospective reverse genetic techniques that are applicable to the zebrafish model. We include discussion of pharmacological approaches, popular gain-of-function and knockdown approaches, and gene targeting strategies. We consider the need for temporal and spatial control over gain/loss of gene function, and discuss available and developing techniques to achieve this end. Our goal is both to reveal the current technical advantages of the zebrafish and to highlight those areas where work is still required to allow this system to be exploited to full advantage.
Skromne
Isaac
I
Department of Biology, University of Miami, Coral Gables, Florida 33146, USA. iskromne@bio.umiami.edu
Prince
Victoria E
VE
D000818:Animals; D030801:Animals, Genetically Modified; D000970:Antineoplastic Agents / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology; D018390:Gene Targeting / Q000379:methods; D005821:Genetic Techniques; D008954:Models, Biological; D016296:Mutagenesis; D016376:Oligonucleotides, Antisense / Q000378:metabolism; D010641:Phenotype; D034622:RNA Interference; D016337:RNA, Catalytic / Q000378:metabolism; D011995:Recombination, Genetic; D014212:Tretinoin / Q000494:pharmacology; D015027:Zebrafish / Q000235:genetics* / Q000254:growth & development; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism
D000970:Antineoplastic Agents; D016376:Oligonucleotides, Antisense; D016337:RNA, Catalytic; D029961:Zebrafish Proteins; D014212:Tretinoin