Dual RMCE for efficient re-engineering of mouse mutant alleles.

Journal:

Nat. Methods 2010 Nov

Authors:

Osterwalder M, Galli A, Rosen B, Skarnes WC, Zeller R, Lopez-Rios J

Abstract

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.

Mesh Headings:

Alleles, Animals, Basic Helix-Loop-Helix Transcription Factors, Carrier Proteins, Embryonic Stem Cells, Genetic Engineering, Mice, Nerve Tissue Proteins, Nuclear Proteins, Promoter Regions, Genetic, Recombinases, Recombination, Genetic, Smad4 Protein