GSBS Student Publications

Title

Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases

UMMS Affiliation

Graduate School of Biomedical Sciences; Program in Gene Function and Expression; Department of Biochemistry and Molecular Pharmacology

Date

5-27-2008

Document Type

Article

Medical Subject Headings

Animals; Animals, Genetically Modified; Deoxyribonucleases; *Gene Silencing; Gene Targeting; Genetic Engineering; Mutagenesis, Site-Directed; Protein Engineering; Zebrafish; Zebrafish Proteins; Zinc Fingers

Disciplines

Life Sciences | Medicine and Health Sciences

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.

Rights and Permissions

Citation: Nat Biotechnol. 2008 Jun;26(6):695-701. Epub 2008 May 25. Link to article on publisher's site

DOI of Published Version

10.1038/nbt1398

Related Resources

Link to Article in PubMed

Journal Title

Nature biotechnology

PubMed ID

18500337