• search hit 10 of 16
Back to Result List

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

  • 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.

Export metadata

Additional Services

Share in Twitter Search Google Scholar Availability
Metadaten
Document Type:Article
Language:English
Parent Title (English):Genesis
Volume:45
Issue:8
First Page:508
Last Page:517
ISSN:1526-954X
DOI:https://doi.org/10.1002/dvg.20321
Pubmed Id:http://www.ncbi.nlm.nih.gov/pubmed?term=17661398
Publisher:Wiley-Blackwell
Date of first publication:2007/07/27
Departments, institutes and facilities:Institut für funktionale Gen-Analytik (IfGA)
Dewey Decimal Classification (DDC):600 Technik, Medizin, angewandte Wissenschaften / 610 Medizin und Gesundheit / 610 Medizin und Gesundheit
Entry in this database:2018/07/14