TY  - JOUR
A1  - Schebelle, Laura
A1  - Wolf, Claudia
A1  - Stribl, Carola
A1  - Javaheri, Tahereh
A1  - Schnütgen, Frank
A1  - Ettinger, Andreas
A1  - Ivics, Zoltán
A1  - Hansen, Jens
A1  - Ruiz, Patricia
A1  - Melchner, Harald von
A1  - Wurst, Wolfgang
A1  - Floss, Thomas
T1  - Efficient conditional and promoter-specific in vivo expression of cDNAs of choice by taking advantage of recombinase-mediated cassette exchange using FlEx gene traps
T2  - Nucleic acids research
N2  - Recombinase-mediated cassette exchange (RMCE) exploits the possibility to unidirectionally exchange any genetic material flanked by heterotypic recombinase recognition sites (RRS) with target sites in the genome. Due to a limited number of available pre-fabricated target sites, RMCE in mouse embryonic stem (ES) cells has not been tapped to its full potential to date. Here, we introduce a universal system, which allows the targeted insertion of any given transcriptional unit into 85 742 previously annotated retroviral conditional gene trap insertions, representing 7013 independent genes in mouse ES cells, by RMCE. This system can be used to express any given cDNA under the control of endogenous trapped promoters in vivo, as well as for the generation of transposon ‘launch pads’ for chromosomal region-specific ‘Sleeping Beauty’ insertional mutagenesis. Moreover, transcription of the gene-of-interest is only activated upon Cre-recombinase activity, a feature that adds conditionality to this expression system, which is demonstrated in vivo. The use of the RMCE system presented in this work requires one single-cloning step followed by one overnight gateway clonase reaction and subsequent cassette exchange in ES cells with efficiencies of 40% in average.
Y1  - 2010
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/20248
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30-85070
SN  - 0305-1048
SN  - 1362-4962
N1  - © The Author(s) 2010. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
VL  - 38
IS  - 9, e106
SP  - 1
EP  - 12
PB  - Oxford Univ. Press
CY  - Oxford
ER  -