Details

Autor(en) / Beteiligte

• Peitz, Michael
• Jäger, Richard
• Patsch, Christoph
• Jäger, Andrea
• Egert, Angela
• Schorle, Hubert
• Edenhofer, Frank

Titel

Enhanced purification of cell-permeant Cre and germline transmission after transduction into mouse embryonic stem cells

Ist Teil von

• Genesis (New York, N.Y. : 2000), 2007-08, Vol.45 (8), p.508-517

Ort / Verlag

Hoboken: Wiley Subscription Services, Inc., A Wiley Company

Erscheinungsjahr

2007

Link zum Volltext

Quelle

Wiley-Blackwell Journals

Beschreibungen/Notizen

• 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. genesis 45:508–517, 2007. © 2007 Wiley‐Liss, Inc.