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A versatile single‐plasmid system for tissue‐specific and inducible control of gene expression in transgenic mice
Author(s) -
Ko JaeKyun,
Choi KyoungHan,
Zhao Xiaoli,
Komazaki Shinji,
Pan Zui,
Weisleder Noah,
Ma Jianjie
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.11-181560
Subject(s) - small hairpin rna , gene knockdown , transgene , biology , microbiology and biotechnology , gene expression , dicer , cre recombinase , rna interference , genetically modified mouse , reporter gene , gene , rna , genetics
We describe a novel transgenic system for tissue‐specific and inducible control of gene expression in mice. The system employs a tetracycline‐responsive CMV promoter that controls transcription of a short‐hairpin RNA (shRNA) that remains nonfunctional until an interrupting reporter cassette is excised by Cre recombinase. Insertion of Dicer and Drosha RNase processing sites within the shRNA allows generation of siRNA to knock down a target gene efficiently. Tissue‐specific shRNA expression is achieved through the use of appropriate inducer mice with tissue‐specific expression of Cre. We applied this system to regulate expression of junctophilins (JPs), genes essential for maintenance of membrane ultrastructure and Ca 2+ signaling in muscle. Transgenic mice with skeletal muscle‐specific expression of shRNA against JP mRNAs displayed no basal change of JP expression before treatment with doxycycline (Dox), while inducible and reversible knockdown of JPs was achieved by feeding mice with Dox‐containing water. Dox‐induced knockdown of JPs led to abnormal junctional membrane structure and Ca 2+ signaling in adult muscle fibers, consistent with essential roles of JPs in muscle development and function. This transgenic approach can be applied for inducible and reversible gene knockdown or gene overexpression in many different tissues, thus providing a versatile system for elucidating the physiological gene function in viable animal models.—Ko, J. ‐K., Choi, K. ‐H., Zhao, X., Komazaki, S., Pan, Z., Weisleder, N., Ma, J. A versatile single‐plasmid system for tissue‐specific and inducible control of gene expression in transgenic mice. FASEB J. 25, 2638–2649 (2011). www.fasebj.org