清脆的
基因组编辑
基因组工程
计算生物学
诱导多能干细胞
基因组
单元格排序
生物
Cas9
合成生物学
转录激活物样效应核酸酶
HEK 293细胞
效应器
计算机科学
遗传学
细胞生物学
胚胎干细胞
细胞
细胞培养
基因
作者
Haribaskar Ramachandran,Soraia Martins,Zacharias Kontarakis,Jean Krutmann,Andrea Rossi
出处
期刊:Life science alliance
[Life Science Alliance]
日期:2021-04-26
卷期号:4 (6): e202101051-e202101051
被引量:48
标识
DOI:10.26508/lsa.202101051
摘要
In the last decade, transcription activator-like effector nucleases and CRISPR-based genome engineering have revolutionized our approach to biology. Because of their high efficiency and ease of use, the development of custom knock-out and knock-in animal or cell models is now within reach for almost every laboratory. Nonetheless, the generation of genetically modified cells often requires a selection step, usually achieved by antibiotics or fluorescent markers. The choice of the selection marker is based on the available laboratory resources, such as cell types, and parameters such as time and cost should also be taken into consideration. Here, we present a new and fast strategy called magnetic-activated genome-edited cell sorting, to select genetically modified cells based on the ability to magnetically sort surface antigens (i.e., tCD19) present in Cas9-positive cells. By using magnetic-activated genome-edited cell sorting, we successfully generated and isolated genetically modified human-induced pluripotent stem cells, primary human fibroblasts, SH-SY5Y neuroblast-like cells, HaCaT and HEK 293T cells. Our strategy expands the genome editing toolbox by offering a fast, cheap, and an easy to use alternative to the available selection methods.
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