Characterization of bacterial artificial chromosome transgenic mice expressing mCherry fluorescent protein substituted for the murine smooth muscle alpha-actin gene.
Characterization of bacterial artificial chromosome transgenic mice expressing mCherry fluorescent protein substituted for the murine smooth muscle alpha-actin gene.
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DOI：
10.1002/dvg.20638
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发表时间：
2010-07
期刊：
影响因子：
1.5
通讯作者：
Hirschi, Karen K.
中科院分区：
文献类型：
--
作者：
Armstrong, John J.;Larina, Irina V.;Dickinson, Mary E.;Zimmer, Warren E.;Hirschi, Karen K.
平滑肌α肌动蛋白（SMA）是一种细胞骨架蛋白，由间充质和平滑肌细胞类型表达，包括壁细胞（血管平滑肌细胞和周细胞）。利用细菌人工染色体（BAC）重组技术，在全长SMA启动子和内含子序列的驱动下，构建了表达膜定位樱桃红荧光蛋白（mCherry）的转基因报告小鼠。我们确定5个独立系的创始人和F1后代含有1-3个mCherry替代BAC载体的拷贝。此外，我们表征了SMA- mcherry在胚胎和成体组织中与内源性SMA的表达关系，发现每个品系的转基因报告基因在所有时间点都再现了内源性SMA的表达。我们还能够使用荧光激活细胞分选（FACS）从胚胎组织中分离出表达SMA的细胞。我们证明该标记可以与其他重要的荧光报告结合，并可用于胚胎心脏动力学的实时成像。因此，这些转基因小鼠将有助于通过FACS分离表达sma的活细胞，并研究sma表达细胞（包括血管平滑肌细胞和周细胞）在胚胎和出生后发育过程中的出现、行为和调控。
Smooth muscle α actin (SMA) is a cytoskeletal protein expressed by mesenchymal and smooth muscle cell types, including mural cells (vascular smooth muscle cells and pericytes). Using Bacterial Artificial Chromosome (BAC) recombineering technology, we generated transgenic reporter mice that express a membrane localized cherry red fluorescent protein (mCherry), driven by the full-length SMA promoter and intronic sequences. We determined that the founders and F1 progeny of five independent lines contain 1-3 copies of the mCherry substituted BAC vector. Furthermore, we characterized the expression of SMA-mCherry in relation to endogenous SMA in the embryo and in adult tissues, and found that the transgenic reporter in each line recapitulated endogenous SMA expression at all time points. We were also able to isolate SMA expressing cells from embryonic tissues using fluorescence activated cell sorting (FACS). We demonstrated that this marker can be combined with other vital fluorescent reporters and it can be used for live imaging of embryonic cardiodynamics. Therefore, these transgenic mice will be useful for isolating live SMA-expressing cells via FACS and for studying the emergence, behavior and regulation of SMA-expressing cells, including vascular smooth muscle cells and pericytes throughout embryonic and postnatal development.
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影响因子：
2.5
作者：
Chandler, Kelly J.;Chandler, Ronald L.;Mortlock, Douglas P.
通讯作者：
Mortlock, Douglas P.
影响因子：
2.5
作者：
MCHUGH, KM
通讯作者：
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影响因子：
48
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通讯作者：
Hyman, Anthony A.
影响因子：
3.5
作者：
Maye, Peter;Stover, Mary Louise;Lichtler, Alexander C.
通讯作者：
Lichtler, Alexander C.
影响因子：
4.4
作者：
Lee, EC;Yu, DG;Copeland, NG
通讯作者：
Copeland, NG