Temporally regulated and tissue-specific gene manipulations in the adult and embryonic heart using a tamoxifen-inducible Cre protein
Temporally regulated and tissue-specific gene manipulations in the adult and embryonic heart using a tamoxifen-inducible Cre protein
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DOI：
10.1161/hh1301.092687
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发表时间：
2001-07-06
影响因子：
20.1
通讯作者：
Molkentin, JD
中科院分区：
文献类型：
--
作者：
Sohal, DS;Nghiem, M;Molkentin, JD
在基因靶向或转基因小鼠中的条件和组织特异性重组系统的出现允许以时间调节和细胞特异性方式评估单基因功能。在这里，我们产生了转基因小鼠表达的他莫昔芬诱导的Cre重组酶蛋白融合到两个突变的雌激素受体配体结合域（MerCreMer）的ar-肌球蛋白重链启动子的控制下。将这些转基因小鼠与ROSA 26 lacZ-flox靶向小鼠杂交，以检查Cre重组酶活性和系统的保真度。数据表明，在没有诱导剂的情况下，胚胎、新生儿或成人心脏中基本上没有Cre介导的重组，但仅注射四次他莫昔芬后就发生了> 80%的重组。MerCreMer融合蛋白在成人心脏内的表达不影响心脏性能、细胞结构或肥大标记基因的表达，表明转基因编码的蛋白相对无害。总之，MerCreMer转基因小鼠代表了一种工具，用于在发育和成年心脏内暂时调节任何loxP靶向基因的失活，或用于特异性指导loxP失活心脏转基因在心脏中的重组和表达。
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 ar-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.