Details

Autor(en) / Beteiligte

• Doyon, Yannick
• Rebar, Edward J
• McCammon, Jasmine M
• Amora, Rainier
• Faraji, Farhoud
• Miller, Jeffrey C
• Gregory, Philip D
• Ngo, Catherine
• Amacher, Sharon L
• Katibah, George E
• Zhang, Lei
• Hocking, Toby D
• Urnov, Fyodor D

Titel

Heritable targeted gene disruption in zebrafish using designed zinc-finger nucleases

Ist Teil von

• Nature biotechnology, 2008-06, Vol.26 (6), p.702-708

Ort / Verlag

New York, NY: Nature Publishing Group

Erscheinungsjahr

2008

Quelle

MEDLINE

Beschreibungen/Notizen

• We describe the use of zinc-finger nucleases (ZFNs) for somatic and germline disruption of genes in zebrafish (Danio rerio), in which targeted mutagenesis was previously intractable. ZFNs induce a targeted double-strand break in the genome that is repaired to generate small insertions and deletions. We designed ZFNs targeting the zebrafish golden and no tail/Brachyury (ntl) genes and developed a budding yeast-based assay to identify the most active ZFNs for use in vivo. Injection of ZFN-encoding mRNA into one-cell embryos yielded a high percentage of animals carrying distinct mutations at the ZFN-specified position and exhibiting expected loss-of-function phenotypes. Over half the ZFN mRNA-injected founder animals transmitted disrupted ntl alleles at frequencies averaging 20%. The frequency and precision of gene-disruption events observed suggest that this approach should be applicable to any loci in zebrafish or in other organisms that allow mRNA delivery into the fertilized egg.