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Nat Biotechnol. 2008 Jun;26(6):695-701. doi: 10.1038/nbt1398. Epub 2008 May 25.

Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases.

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  • 1Program in Gene Function and Expression, University of Massachusetts Medical School, Lazare Research Building, 6th Floor, 364 Plantation St., Worcester, Massachusetts 01605, USA.

Abstract

Direct genomic manipulation at a specific locus is still not feasible in most vertebrate model organisms. In vertebrate cell lines, genomic lesions at a specific site have been introduced using zinc-finger nucleases (ZFNs). Here we adapt this technology to create targeted mutations in the zebrafish germ line. ZFNs were engineered that recognize sequences in the zebrafish ortholog of the vascular endothelial growth factor-2 receptor, kdr (also known as kdra). Co-injection of mRNAs encoding these ZFNs into one-cell-stage zebrafish embryos led to mutagenic lesions at the target site that were transmitted through the germ line with high frequency. The use of engineered ZFNs to introduce heritable mutations into a genome obviates the need for embryonic stem cell lines and should be applicable to most animal species for which early-stage embryos are easily accessible.

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PMID:
18500337
[PubMed - indexed for MEDLINE]
PMCID:
PMC2502069
Free PMC Article

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