Journal
DEVELOPMENTAL DYNAMICS
Volume 237, Issue 4, Pages 861-882Publisher
WILEY
DOI: 10.1002/dvdy.21484
Keywords
reverse genetics; pharmacology; mRNA; over-expression; dominant negative; gain-of-function; loss-offunction; temporal regulation; caged RNA; heat shock; antisense; morpholino; peptide nucleic acid; ribozyme; RNA interference; siRNA; miRNA; dsRNA; transgenesis; retrovirus; transposons; Tol2; Cre/lox; Tet-on; Gal4/UAS; insertional mutagenesis; target gene inactivation; homologous recombination; target-selected mutagenesis; TILLING; retinoic acid
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Funding
- NIDDK NIH HHS [DK064973-01] Funding Source: Medline
- NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES [R01DK064973, R56DK064973] Funding Source: NIH RePORTER
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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.
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