Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases
- Meng, X., Noyes, M.B., Zhu, L.J., Lawson, N.D., and Wolfe, S.A.
- Nat. Biotechnol. 26(6): 695-701 (Journal)
- Registered Authors
- Lawson, Nathan, Wolfe, Scot A.
- MeSH Terms
- Animals, Genetically Modified/physiology*
- Gene Silencing*
- Gene Targeting/methods*
- Genetic Engineering/methods*
- Mutagenesis, Site-Directed/methods
- Protein Engineering/methods
- Zebrafish Proteins/genetics*
- Zinc Fingers/genetics*
- 18500337 Full text @ Nat. Biotechnol.
Meng, X., Noyes, M.B., Zhu, L.J., Lawson, N.D., and Wolfe, S.A. (2008) Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases. Nat. Biotechnol.. 26(6):695-701.
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.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes