TY  - JOUR
A1  - Peitz, Michael
A1  - Jäger, Richard
A1  - Patsch, Christoph
A1  - Jäger, Andrea
A1  - Egert, Angela
A1  - Schorle, Hubert
A1  - Edenhofer, Frank
T1  - Enhanced purification of cell-permeant Cre and germline transmission after transduction into mouse embryonic stem cells
T2  - Genesis
N2  - Continuous expression of Cre recombinase has the potential to yield toxic side effects in various cell types, thereby limiting applications of the Cre/loxP system for conditional mutagenesis. In this study, we investigate the potential of Cre protein transduction to overcome this limitation. COS-7, CV1-5B, and mouse embryonic stem (ES) cells treated with cell-permeant Cre (HTNCre) maintain a normal growth behavior employing Cre concentrations sufficient to induce recombination in more than 90% of the cells, whereas continuous application of high doses resulted in markedly reduced proliferation. HTNCre-treated ES cells maintain a normal karyotype and are still able to contribute to the germline. Moreover, we present an enhanced HTNCre purification protocol that allows the preparation of a concentrated glycerol stock solution, thereby enabling a considerable simplification of the Cre protein transduction procedure. The protocol described here allows rapid and highly efficient conditional mutagenesis of cultured cells.
Y1  - 2007
UR  - https://pub.h-brs.de/frontdoor/index/index/docId/3751
SN  - 1526-954X
VL  - 45
IS  - 8
SP  - 508
EP  - 517
PB  - Wiley-Blackwell
ER  -