Abstract
Animal models with genetic modifications under temporal and/or spatial control are invaluable to functional genomics and medical research. Here we report the generation of tissue-specific knockout rats via microinjection of zinc-finger nucleases (ZFNs) into fertilized eggs. We generated rats with loxP-flanked (floxed) alleles and a tyrosine hydroxylase promoter–driven cre allele and demonstrated Cre-dependent gene disruption in vivo. Pronuclear microinjection of ZFNs, shown by our data to be an efficient and rapid method for creating conditional knockout rats, should also be applicable in other species.
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26 August 2013
In the version of this article initially published, a name was misspelled in the Acknowledgements section. The error has been corrected in the HTML and PDF versions of the article.
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Acknowledgements
We thank A. Lambowitz for critical advice, F. Urnov, D. Carroll and G. Davis for valuable comments on the manuscript, J. Huang (Cold Spring Harbor Lab) for providing IRES-cre sequence, K. Watsek for animal husbandry, L. Little for technical assistance and G. Zhao for suggestions.
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X.C. designed the experiments and analyzed the data; A.J.B. and D.A.F. carried out experiments; A.J.B., Y.W. and J.W. did microinjections; R.H., D.J., A.C. and W.S. did part of the genotyping; E.K. and K.F. performed qRT-PCR; A.M. did western blots; and X.C. and E.J.W. wrote the manuscript.
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All authors are full-time employees of SAGE Labs, Inc., which sells genetically engineered rats and provides custom model creation services.
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Brown, A., Fisher, D., Kouranova, E. et al. Whole-rat conditional gene knockout via genome editing. Nat Methods 10, 638–640 (2013). https://doi.org/10.1038/nmeth.2516
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DOI: https://doi.org/10.1038/nmeth.2516
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