敲除（某基因）会导致大鼠早期胚胎死亡。 （你原文中“Knockout of ”后面缺少具体基因信息）
Knockout of causes early embryonic death in rats.
作者信息
Yang Xinlan, Lu Dan, Zhang Xu, Chen Wei, Gao Shan, Dong Wei, Ma Yuanwu, Zhang Lianfeng
机构信息
Key Laboratory of Human Disease Comparative Medicine National Health Commission of China (NHC) Institute of Laboratory Animal Science Peking Union Medical College Chinese Academy of Medical Sciences Beijing China.
Beijing Engineering Research Center for Experimental Animal Models of Human Diseases Institute of Laboratory Animal Science Peking Union Medical College Chinese Academy of Medical Sciences Beijing China.
出版信息
Animal Model Exp Med. 2019 Mar 12;2(1):18-24. doi: 10.1002/ame2.12059. eCollection 2019 Mar.
BACKGROUND
Iron-sulfur cluster assembly 1 (ISCA1) is an iron-sulfur (Fe/S) carrier protein that accepts Fe/S from a scaffold protein and transfers it to target proteins including the mitochondrial Fe/S containing proteins. is also the newly identified causal gene for multiple mitochondrial dysfunctions syndrome (MMDS). However, our knowledge about the physiological function of ISCA1 in vivo is currently limited. In this study, we generated an knockout rat line and analyzed the embryo development.
METHODS
knockout rats were generated by replacing the exon1 of gene with the fusion gene using CRISPR-Cas9 technology. The ISCA1 expression pattern was analyzed by fluorescence imaging using ISCA1 promotor driven Cre and mCherry expression. The embryonic morphology was examinated by microscope and mitochondrial proteins were tested by Western blot.
RESULTS
An knockout rat line was obtained, which expressed mCherry-Cre fusion protein. Both of the fluorescence images from mCherry and Cre induced mCherry in a reporter rat strain, showing that ISCA1 expressed in most of the tissues in rats. The ISCA1 knockout resulted in abnormal development at 8.5 days, with a significant decrease of NDUFA9 protein and an increase of aconitase 2 (ACO2) in rat embryos.
CONCLUSION
Deletion of induced early death in rats. ISCA1 affected the expression of key proteins in the mitochondrial respiratory chain complex, suggesting that ISCA1 has an important influence on the respiratory complex and energy metabolism.
背景
铁硫簇组装蛋白1（ISCA1）是一种铁硫（Fe/S）载体蛋白，它从支架蛋白接受Fe/S并将其转移到目标蛋白，包括含线粒体Fe/S的蛋白。它也是新发现的导致多种线粒体功能障碍综合征（MMDS）的致病基因。然而，我们目前对ISCA1在体内生理功能的了解有限。在本研究中，我们构建了ISCA1基因敲除大鼠品系并分析胚胎发育情况。
方法
使用CRISPR-Cas9技术，用mCherry-Cre融合基因替换ISCA1基因的外显子1来构建ISCA1基因敲除大鼠。通过使用ISCA1启动子驱动的Cre和mCherry表达的荧光成像分析ISCA1的表达模式。用显微镜检查胚胎形态，并用蛋白质免疫印迹法检测线粒体蛋白。
结果
获得了一个ISCA1基因敲除大鼠品系，其表达mCherry-Cre融合蛋白。来自mCherry的荧光图像以及在报告基因大鼠品系中由Cre诱导的mCherry荧光图像，均表明ISCA1在大鼠的大多数组织中表达。ISCA1基因敲除导致大鼠胚胎在8.5天时发育异常，大鼠胚胎中NDUFA9蛋白显著减少，乌头酸酶2（ACO2）增加。
结论
ISCA1基因缺失导致大鼠早期死亡。ISCA1影响线粒体呼吸链复合物关键蛋白的表达，提示ISCA1对呼吸复合物和能量代谢有重要影响。
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