Open AccessGerm-Line Recombination Activity of the Widely Used hGFAP-Cre and Nestin-Cre Transgenes
Author(s) -
Jiong Zhang,
Pavel Dublin,
Stephanie Griemsmann,
Annette Klein,
Ralph Brehm,
Peter Bedner,
Bernd K. Fleischmann,
Christian Steinhaüser,
Martin Theis
Publication year - 2013
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0082818
Subject(s) - transgene , biology , genetically modified mouse , nestin , microbiology and biotechnology , germline , gene , glial fibrillary acidic protein , gene targeting , genetics , neural stem cell , stem cell , immunohistochemistry , immunology
Herein we demonstrate with PCR, immunodetection and reporter gene approaches that the widely used human Glial Fibrillary Acidic Protein (hGFAP)-Cre transgene exhibits spontaneous germ-line recombination activity in leading to deletion in brain, heart and tail tissue with high frequency. The ectopic activity of hGFAP-Cre requires a rigorous control. We likewise observed that a second widely used nestin-Cre transgene shows germ-line deletion. Here we describe procedures to identify mice with germ-line recombination mediated by the hGFAP-Cre and nestin-Cre transgenes. Such control is essential to avoid pleiotropic effects due to germ-line deletion of loxP-flanked target genes and to maintain the CNS-restricted deletion status in transgenic mouse colonies.