Whole-rat conditional gene knockout via genome editing

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  • A Corrigendum to this article was published on 27 September 2013

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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Figure 1: Floxing Grin1 exon 4 using the two-cut strategy.

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NCBI Reference Sequence

Change history

  • 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.

Author information

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.

Correspondence to Xiaoxia Cui.

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Competing interests

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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Supplementary Figures 1–10, Supplementary Tables 1–5 and Supplementary Note (PDF 1425 kb)

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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) doi:10.1038/nmeth.2516

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