Brief Communication | Published:

Whole-rat conditional gene knockout via genome editing

Nature Methods volume 10, pages 638640 (2013) | Download Citation

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

This article has been updated

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.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

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.

Accessions

References

  1. 1.

    et al. Science 325, 433 (2009).

  2. 2.

    et al. Nat. Biotechnol. 29, 64–67 (2011).

  3. 3.

    et al. Nat. Biotechnol. 29, 695–696 (2011).

  4. 4.

    , , , & Science 265, 103–106 (1994).

  5. 5.

    et al. Nat. Methods 8, 753–755 (2011).

  6. 6.

    , , , & Dis. Model. Mech. 6, 269–278 (2013).

  7. 7.

    et al. Neuron 71, 995–1013 (2011).

  8. 8.

    et al. Neuron 13, 325–338 (1994).

  9. 9.

    , & Nature 374, 640–643 (1995).

  10. 10.

    & Int. J. Biochem. Cell Biol. 37, 942–946 (2005).

  11. 11.

    et al. Neuron 72, 721–733 (2011).

  12. 12.

    & in Biology of the NMDA Receptor (ed. Van Dongen, A.M.) Ch. 5 (CRC Press, 2009).

  13. 13.

    , , & PLoS ONE 6, e28283 (2011).

  14. 14.

    et al. Biochem. Biophys. Res. Commun. 319, 1197–1202 (2004).

  15. 15.

    Nature 372, 143–149 (1994).

  16. 16.

    et al. Nat. Genet. 22, 375–378 (1999).

  17. 17.

    , & Nat. Genet. 37, 1187–1193 (2005).

Download references

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

Affiliations

  1. Sage Labs, St. Louis, Missouri, USA.

    • Andrew J Brown
    • , Daniel A Fisher
    • , Evguenia Kouranova
    • , Aaron McCoy
    • , Kevin Forbes
    • , Yumei Wu
    • , Rachel Henry
    • , Diana Ji
    • , Andre Chambers
    • , Joe Warren
    • , Weiguo Shu
    • , Edward J Weinstein
    •  & Xiaoxia Cui

Authors

  1. Search for Andrew J Brown in:

  2. Search for Daniel A Fisher in:

  3. Search for Evguenia Kouranova in:

  4. Search for Aaron McCoy in:

  5. Search for Kevin Forbes in:

  6. Search for Yumei Wu in:

  7. Search for Rachel Henry in:

  8. Search for Diana Ji in:

  9. Search for Andre Chambers in:

  10. Search for Joe Warren in:

  11. Search for Weiguo Shu in:

  12. Search for Edward J Weinstein in:

  13. Search for Xiaoxia Cui in:

Contributions

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.

Competing interests

All authors are full-time employees of SAGE Labs, Inc., which sells genetically engineered rats and provides custom model creation services.

Corresponding author

Correspondence to Xiaoxia Cui.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–10, Supplementary Tables 1–5 and Supplementary Note

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nmeth.2516

Further reading

Newsletter Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing