Smooth muscle expression of Cre recombinase and eGFP in transgenic mice
Smooth muscle expression of Cre recombinase and eGFP in transgenic mice
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DOI：
10.1152/physiolgenomics.00054.2002
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发表时间：
2002-09-03
影响因子：
4.6
通讯作者：
Kotlikoff, MI
中科院分区：
文献类型：
--
作者：
Xin, HB;Deng, KY;Kotlikoff, MI
转基因小鼠中Cre重组酶和eGFP的平滑肌表达。Physiol Genomics 10：211- 215，2002.首次发表于2002年7月2日; 10.1152/ physiolgenomics。00054.2002.-我们报告了转基因小鼠的产生，旨在促进血管和非血管平滑肌生物学在体内的研究。平滑肌肌球蛋白重链（smMHC）启动子用于指导由Cre重组酶和增强型绿色荧光蛋白（eGFP）编码序列组成的双顺反子转基因的表达。表达转基因的动物显示出局限于血管和非血管平滑肌的强荧光。平滑肌的酶促解离产生活的荧光细胞，其可以作为单细胞进行研究或通过FACS分选用于基因表达研究。将smMHC/ Cre/ eGFP小鼠与ROSA 26/ lacZ报告小鼠杂交以确定Cre重组酶活性; Cre重组酶在成年小鼠的所有平滑肌中表达，并且重组酶和eGFP的表达之间存在极好的重叠。荧光和Cre重组酶在胚胎第12.5天检测到平滑肌特异性表达.这些小鼠将是有用的，以确定在小鼠体内的平滑肌基因功能，在单个活细胞中的基因功能的研究，并在血管和非血管平滑肌中的基因表达的测定。
Smooth muscle expression of Cre recombinase and eGFP in transgenic mice. Physiol Genomics 10: 211- 215, 2002. First published July 2, 2002; 10.1152/ physiolgenomics. 00054.2002.- We report the generation of transgenic mice designed to facilitate the study of vascular and nonvascular smooth muscle biology in vivo. The smooth muscle myosin heavy chain (smMHC) promoter was used to direct expression of a bicistronic transgene consisting of Cre recombinase and enhanced green fluorescent protein (eGFP) coding sequences. Animals expressing the transgene display strong fluorescence confined to vascular and nonvascular smooth muscle. Enzymatic dissociation of smooth muscle yields viable, fluorescent cells that can be studied as single cells or sorted by FACS for gene expression studies. smMHC/ Cre/ eGFP mice were crossed with ROSA26/ lacZ reporter mice to determine Cre recombinase activity; Cre recombinase was expressed in all smooth muscles in adult mice, and there was an excellent overlap between expression of the recombinase and eGFP. Initial smooth muscle- specific expression of fluorescence and Cre recombinase was detected on embryonic day 12.5. These mice will be useful to define smooth muscle gene function in vivo in mice, for the study of gene function in single, live cells, and for the determination of gene expression in vascular and nonvascular smooth muscle.