Temporally Regulated and Tissue-Specific Gene Manipulations in the Adult and Embryonic Heart Using a Tamoxifen-Inducible Cre Protein

TLDR

Conditional, tissue‑specific recombination systems in mice enable temporally regulated, cell‑specific assessment of single gene function. The authors generated α‑MHC‑driven MerCreMer transgenic mice and crossed them with ROSA26 lacZ‑flox mice to evaluate Cre activity and system fidelity. The MerCreMer mice showed no Cre activity without tamoxifen, >80 % recombination after four injections, and did not alter cardiac performance, making them a safe, temporally regulated tool for heart‑specific gene manipulation.

Abstract

Abstract —The advent of conditional and tissue-specific recombination systems in gene-targeted or transgenic mice has permitted an assessment of single gene function in a temporally regulated and cell-specific manner. Here we generated transgenic mice expressing a tamoxifen-inducible Cre recombinase protein fused to two mutant estrogen-receptor ligand-binding domains (MerCreMer) under the control of the α-myosin heavy chain promoter. These transgenic mice were crossed with the ROSA26 lacZ-flox –targeted mice to examine Cre recombinase activity and the fidelity of the system. The data demonstrate essentially no Cre-mediated recombination in the embryonic, neonatal, or adult heart in the absence of inducing agent but >80% recombination after only four tamoxifen injections. Expression of the MerCreMer fusion protein within the adult heart did not affect cardiac performance, cellular architecture, or expression of hypertrophic marker genes, demonstrating that the transgene-encoded protein is relatively innocuous. In summary, MerCreMer transgenic mice represent a tool for temporally regulated inactivation of any loxP-targeted gene within the developing and adult heart or for specifically directing recombination and expression of a loxP-inactivated cardiac transgene in the heart.