Journal
NATURE BIOTECHNOLOGY
Volume 26, Issue 6, Pages 695-701Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nbt1398
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Funding
- NHLBI NIH HHS [R01 HL079266, R01 HL079266-05, R01HL079266] Funding Source: Medline
- NIGMS NIH HHS [R01 GM068110, 1R01GM068110, R01 GM068110-05] Funding Source: Medline
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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)(1-3). 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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