Open AccessTissue‐Specific Regulation of Oncogene Expression Using Cre‐Inducible ROSA26 Knock‐In Transgenic Mice
Author(s) -
Carofino Brandi L.,
Justice Monica J.
Publication year - 2015
Publication title -
current protocols in mouse biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.77
H-Index - 15
ISSN - 2161-2617
DOI - 10.1002/9780470942390.mo140150
Subject(s) - cre recombinase , gene knockin , transgene , biology , recombinase , genetically modified mouse , germline , transgenesis , cre lox recombination , gene targeting , microbiology and biotechnology , gene , genetics , embryogenesis , reproductive biology , recombination
Cre‐inducible mouse models are often utilized for the spatial and temporal expression of oncogenes. With the wide number of Cre recombinase lines available, inducible transgenesis represents a tractable approach to achieve discrete oncogene expression. Here, we describe a protocol for targeting Cre‐inducible genes to the ubiquitously expressed ROSA26 locus. Gene targeting provides several advantages over standard transgenic techniques, including a known site of integration and previously characterized pattern of expression. Historically, an inherent instability of ROSA26 targeting vectors has hampered the efficiency of developing ROSA26 knock‐in lines. In this protocol, we provide individual steps for utilizing Gateway recombination for cloning as well as detailed instructions for screening targeted ES cell clones. By following this protocol, one can achieve germline transmission of a ROSA26 knock‐in line within several months. © 2015 by John Wiley & Sons, Inc.