利用CRISPR/Cas9系统生成CLTA报告基因人诱导多能干细胞系CRMi001-A-1，以监测内源性网格蛋白运输。
Generation of a CLTA reporter human induced pluripotent stem cell line, CRMi001-A-1, using the CRISPR/Cas9 system to monitor endogenous clathrin trafficking.
作者信息
Anderson Ruthellen H, Kerkvliet Jason G, Otta Jaelin J, Ross Alan D, Leiferman Patricia C, Hoppe Adam D, Francis Kevin R
机构信息
Sanford School of Medicine, University of South Dakota, Sioux Falls, SD, USA; Cellular Therapies and Stem Cell Biology Group, Sanford Research, Sioux Falls, SD, USA.
Department of Chemistry and Biochemistry, South Dakota State University, Brookings, SD, USA; BioSystems Networks and Translational Research Center, Brookings, SD, USA.
出版信息
Stem Cell Res. 2018 Dec;33:95-99. doi: 10.1016/j.scr.2018.10.001. Epub 2018 Oct 3.
The most highly studied endocytic pathway, clathrin-dependent endocytosis, mediates a wide range of fundamental processes including nutrient internalization, receptor recycling, and signal transduction. In order to model tissue specific and developmental aspects of this process, CRISPR/Cas9 genomic editing was utilized to fluorescently label the C-terminus of clathrin light chain A (CLTA) within the phenotypically normal, parental CRMi001-A human induced pluripotent stem cell line. Successfully edited cells were isolated by fluorescently activated cell sorting, remained karyotypically normal, and maintained their differentiation potential. This cell line facilitates imaging of endogenous clathrin trafficking within varied cell types.
研究最为深入的内吞途径——网格蛋白依赖的内吞作用，介导了包括营养物质内化、受体循环利用和信号转导在内的一系列重要过程。为了模拟这一过程的组织特异性和发育方面，利用CRISPR/Cas9基因组编辑技术，在表型正常的亲本CRMi001-A人诱导多能干细胞系中对网格蛋白轻链A（CLTA）的C末端进行荧光标记。通过荧光激活细胞分选分离出成功编辑的细胞，这些细胞的核型保持正常，并维持其分化潜能。该细胞系有助于对内源网格蛋白在不同细胞类型中的运输进行成像。
Zotero 插件