PUBLICATION
Heritable targeted gene disruption in zebrafish using designed zinc-finger nucleases
- Authors
- Doyon, Y., McCammon, J.M., Miller, J.C., Faraji, F., Ngo, C., Katibah, G.E., Amora, R., Hocking, T.D., Zhang, L., Rebar, E.J., Gregory, P.D., Urnov, F.D., and Amacher, S.L.
- ID
- ZDB-PUB-080527-24
- Date
- 2008
- Source
- Nat. Biotechnol. 26(6): 702-708 (Journal)
- Registered Authors
- Amacher, Sharon, McCammon, Jasmine
- Keywords
- none
- MeSH Terms
-
- Deoxyribonucleases/genetics
- Animals, Genetically Modified/physiology*
- Animals
- Gene Targeting/methods*
- Protein Engineering/methods
- PubMed
- 18500334 Full text @ Nat. Biotechnol.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping