Generation of MLC-2v-tdTomato knock-in reporter mouse line.

Generation of MLC-2v-tdTomato knock-in reporter mouse line.
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DOI:
10.1002/dvg.23256
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发表时间:
2018-10
期刊:
Genesis (New York, N.Y. : 2000)
影响因子:
--
通讯作者:
Nam YJ
Nam YJ
中科院分区:
其他
文献类型:
--
作者:
Zhang Z;Nam YJ

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MLC-2v是一种肌球蛋白轻链调节蛋白,在心室心肌细胞和慢抽搐骨骼肌细胞中特异性表达。在心脏发育过程中,MLC-2v在心室成熟中起关键作用。缺乏MLC-2v的小鼠由于心室心肌细胞肌纤维组织异常导致心力衰竭而致胚胎死亡。为了研究室性心肌和慢抽动骨骼肌的发育,我们在不破坏内源基因的情况下,将tdTomato报告盒敲入MLC-2v基因的3 ' UTR,产生了新的MLC-2v报告小鼠系。我们的研究结果表明,在心肌发生过程中,MLC-2v- tdtomato敲入报告基因在心室心肌细胞和慢抽动肌中有特异性表达,准确地概括了内源性MLC-2v的时空表达模式。在整个发育过程中,心房、快速收缩肌或其他器官均未观察到tdTomato的表达。分离的新生儿和成人心室心肌细胞一致表达tdTomato。综上所述,本文所描述的MLC-2v-tdTomato敲入报告小鼠模型将克服转基因策略的缺陷,为研究心脏室和骨骼肌在发育和再生过程中的特征提供有价值的工具。
MLC-2v is a myosin light chain regulatory protein which is specifically expressed in ventricular cardiomyocytes and slow twitch skeletal muscle cells. MLC-2v plays critical roles in ventricular maturation during heart development. Mice lacking MLC-2v are embryonic lethal due to heart failure associated with abnormal myofibrillar organization of ventricular cardiomyocytes. To study the development of ventricular cardiac muscle and slow twitch skeletal muscle, we generated a new MLC-2v reporter mouse line by knocking-in a tdTomato reporter cassette into 3’ UTR of the MLC-2v gene without disrupting the endogenous gene. Our results demonstrated specific MLC-2v-tdTomato knock-in reporter expression in ventricular cardiomyocytes and slow twitch muscle during myogenesis, precisely recapitulating the spatiotemporal expression pattern of endogenous MLC-2v. No tdTomato expression was observed in the atria, fast twitch muscle or other organs throughout development into adulthood. Isolated neonatal and adult ventricular cardiomyocytes uniformly express tdTomato. Taken together, MLC-2v-tdTomato knock-in reporter mouse model described in this article will serve as a valuable tool to study cardiac chamber and skeletal muscle specification during development and regeneration by overcoming the pitfalls of transgenic strategies.
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