诱导多能干细胞
胚胎干细胞
生物
基因靶向
同源盒蛋白纳米
腺相关病毒
病毒载体
电穿孔
分子生物学
同源重组
干细胞
遗传学
细胞生物学
基因
载体(分子生物学)
重组DNA
作者
K Mitsui,Keiichiro Suzuki,Emi Aizawa,Eihachiro Kawase,Hirofumi Suemori,Norio Nakatsuji,Kohnosuke Mitani
标识
DOI:10.1016/j.bbrc.2009.08.075
摘要
Human pluripotent stem cells, such as embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs), have the ability to differentiate into various cell types, and will become a potential source of cellular materials for regenerative medicine. To make full use of hESCs or hiPSCs for both basic and clinical research, genetic modification, especially gene targeting via homologous recombination (HR), would be an essential technique. This report describes the successful gene targeting of the hypoxanthine phosphoribosyl transferase 1 (HPRT1) and the NANOG loci in human pluripotent stem cells with adeno-associated virus (AAV) vectors. At the HPRT1 locus, up to 1% of stable transformants were targeted via HR with an AAV-HPRT1 targeting vector, without loss of pluripotency. On the other hand, 20–87% of stable transformants were targeted using an AAV-NANOG-targeting vector designed for the promoter-trap strategy. In the KhES-3 cell line, which shows particularly high fragility to experimental manipulation, gene targeting was successful only by using an AAV vector but not by electroporation. In addition to hESC, gene targeting was achieved in hiPSC lines at similar frequencies. These data indicate that AAV vectors may therefore be a useful tool to introduce genetic modifications in hESCs and hiPSCs.
科研通智能强力驱动
Strongly Powered by AbleSci AI