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Genome Editing with CompoZr Custom Zinc Finger Nucleases (ZFNs)
Published on: June 14, 2012
Heritable targeted gene disruption in zebrafish using designed zinc-finger nucleases
Yannick Doyon1, Jasmine M McCammon, Jeffrey C Miller
1Sangamo BioSciences, Inc., Pt. Richmond Tech Center, 501 Canal Blvd., Suite A100, Richmond, California 94804, USA.
Nature Biotechnology
|May 27, 2008
Summary
Zinc-finger nucleases (ZFNs) enable precise gene editing in zebrafish, overcoming previous limitations in targeted mutagenesis. This powerful technique facilitates efficient gene disruption in somatic and germline cells for research applications.
Area of Science:
- Molecular Biology
- Genetics
- Developmental Biology
Background:
- Targeted gene modification in zebrafish (Danio rerio) has historically been challenging.
- Zinc-finger nucleases (ZFNs) offer a potential solution for precise genome engineering.
Purpose of the Study:
- To establish and validate the use of ZFNs for efficient gene disruption in zebrafish.
- To demonstrate the applicability of ZFNs for both somatic and germline gene editing.
Main Methods:
- Design and validation of ZFNs targeting specific zebrafish genes (golden and no tail/Brachyury).
- Utilized a budding yeast-based assay to screen for highly active ZFNs.
- Injected ZFN-encoding mRNA into one-cell zebrafish embryos.
Main Results:
- High efficiency of targeted gene disruption observed in founder zebrafish.
- Generated animals exhibited expected loss-of-function phenotypes.
- Successful transmission of disrupted alleles to offspring with an average frequency of 20%.
Conclusions:
- ZFNs provide a robust and precise method for targeted gene disruption in zebrafish.
- This approach is adaptable for various genes and potentially other organisms with similar embryonic development.
