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Strong and ubiquitous expression of transgenes targeted into the β‐actin locus by Cre/lox cassette replacement
Author(s) -
Shmerling Doron,
Danzer ClausPeter,
Mao Xiaohong,
Boisclair Julie,
Haffner Michel,
Lemaistre Marianne,
Schuler Valerie,
Kaeslin Edgar,
Korn Reinhard,
Bürki Kurt,
Ledermann Birgit,
Kinzel Bernd,
Müller Matthias
Publication year - 2005
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/gene.20135
Subject(s) - transgenesis , transgene , biology , cre recombinase , expression cassette , green fluorescent protein , embryonic stem cell , recombinase , genetically modified mouse , microbiology and biotechnology , endogeny , context (archaeology) , genetics , gene , embryogenesis , recombinant dna , reproductive technology , vector (molecular biology) , biochemistry , recombination , paleontology
Conventional approaches to produce transgenic mice recurrently yield unpredictable patterns and levels of transgene expression, a situation calling for the development of new techniques to overcome these drawbacks in the context of overexpression studies. Here we present an efficient method for rapid and reproducible transgenesis using the recombinase mediated cassette exchange (RMCE) (Bouhassira et al. : Blood 90:3332–3344, 1997) procedure. A lox511‐EGFP‐TK/neo‐loxP cassette was placed under the control of the endogenous mouse β‐actin promoter. Heterozygous mice revealed strong and ubiquitous EGFP expression throughout embryogenesis and adulthood. Reproducibly, the same expression pattern was obtained with RMCE when it was used to replace the EGFP‐harboring cassette by ECFP or placental alkaline phosphatase (PLAP) reporter genes (DePrimo et al. : Transgenic Res 5:459–466, 1996). Furthermore, the RMCE procedure proved efficient as well in embryonic stem (ES) cells as directly in zygotes. Our results demonstrate ubiquitous expression of floxed transgenes in the endogenous β‐actin locus and they support the general use of the β‐actin locus for targeted transgenesis. genesis 42:229–235, 2005. © 2005 Wiley‐Liss, Inc.