Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Dual RMCE for efficient re-engineering of mouse mutant alleles


We have developed dual recombinase-mediated cassette exchange (dRMCE) to efficiently re-engineer the thousands of available conditional alleles in mouse embryonic stem cells. dRMCE takes advantage of the wild-type loxP and FRT sites present in these conditional alleles and in many gene-trap lines. dRMCE is a scalable, flexible tool to introduce tags, reporters and mutant coding regions into an endogenous locus of interest in an easy and highly efficient manner.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: The principle of dRMCE to re-engineer mouse conditional alleles.
Figure 2: dRMCE for efficient modification of difficult-to-target loci.


  1. Glaser, S., Anastassiadis, K. & Stewart, A.F. Nat. Genet. 37, 1187–1193 (2005).

    CAS  Article  Google Scholar 

  2. Branda, C.S. & Dymecki, S.M. Dev. Cell 6, 7–28 (2004).

    CAS  Article  Google Scholar 

  3. Lauth, M., Spreafico, F., Dethleffsen, K. & Meyer, M. Nucleic Acids Res. 30, e115 (2002).

    Article  Google Scholar 

  4. Collins, F.S., Rossant, J. & Wurst, W. Cell 128, 9–13 (2007).

    CAS  Article  Google Scholar 

  5. Shimshek, D.R. et al. Genesis 32, 19–26 (2002).

    CAS  Article  Google Scholar 

  6. Raymond, C.S. & Soriano, P. PLoS ONE 2, e162 (2007).

    Article  Google Scholar 

  7. Kanda, T., Sullivan, K.F. & Wahl, G.M. Curr. Biol. 8, 377–385 (1998).

    CAS  Article  Google Scholar 

  8. Galli, A. et al. PLoS Genet. 6, e1000901 (2010).

    Article  Google Scholar 

  9. Nagy, A. Manipulating the Mouse Embryo: A Laboratory Manual 3rd edn. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA, 2003).

  10. Pettitt, S.J. et al. Nat. Methods 6, 493–495 (2009).

    CAS  Article  Google Scholar 

  11. Anastassiadis, K. et al. Dis. Model, Mech. 2, 508–515 (2009).

    CAS  Article  Google Scholar 

  12. Singla, V. et al. Nat. Methods 7, 50–52 (2010).

    CAS  Article  Google Scholar 

  13. Skarnes, W.C. et al. Nat. Genet. 36, 543–544 (2004).

    CAS  Article  Google Scholar 

  14. Szymczak, A.L. et al. Nat. Biotechnol. 22, 589–594 (2004).

    CAS  Article  Google Scholar 

  15. Chu, G.C., Dunn, N.R., Anderson, D.C., Oxburgh, L. & Robertson, E.J. Development 131, 3501–3512 (2004).

    CAS  Article  Google Scholar 

Download references


We are grateful to D. Klewe-Nebenius and T. Hennek (University of Basel) for generating the chimeric mice. We thank I. Verma (Salk Institute, San Diego, California, USA), B. Sauer (Stowers Institute, Kansas City, Missouri, USA), R. Jaenisch (Whitehead Institute, Cambridge, Massachusetts, USA) and P. Soriano (Mount Sinai School of Medicine, New York, New York, USA) for plasmids obtained via Addgene and A.-K. Hadjantonakis (Sloan-Kettering Institute, New York, New York, USA) for suggesting the use of the H2B-Venus fusion protein. We thank A. Schauerte and P. Lorentz for expert technical assistance and are indebted to P. Bovolenta, G. Nusspaumer, A. Zuniga and members of our research groups for helpful input and discussions. Our research is supported by the Swiss National Science Foundation (to R.Z.), by a Marie Curie Intra-European Fellowship and a European Reintegration Grant (to J.L.-R.) and by both cantons of Basel, Switzerland.

Author information

Authors and Affiliations



M.O., J.L.-R. and R.Z. conceived and designed the experiments. M.O., J.L.-R. and B.R. designed and constructed the dRMCE tool-kit vectors. B.R. and W.C.S. provided the IKMC mouse embryonic stem cell lines. M.O., A.G., J.L.-R. and B.R. performed the experiments. J.L.-R., W.C.S. and R.Z. wrote the paper.

Corresponding authors

Correspondence to Rolf Zeller or Javier Lopez-Rios.

Ethics declarations

Competing interests

A dRMCE patent is pending.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–5, Supplementary Tables 1 and 2 (PDF 8023 kb)

Supplementary Data

Vector sequences in GenBank format. (ZIP 30 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Osterwalder, M., Galli, A., Rosen, B. et al. Dual RMCE for efficient re-engineering of mouse mutant alleles. Nat Methods 7, 893–895 (2010).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

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