Oka Masahiro, Chang Lung-Ji, Costantini Frank, Terada Naohiro
Department of Pathology, University of Florida College of Medicine, Gainesville, FL, USA.
Methods Mol Biol. 2006;329:273-81. doi: 10.1385/1-59745-037-5:273.
The major limitations in gene transduction to embryonic stem (ES) cells are (1) low efficiency of gene delivery and (2) suppression of gene expression after integration into the host genome. A human immunodeficiency virus type I (HIV-1)-based lentiviral vector has been demonstrated to be an excellent tool for stable and efficient gene expression in ES cells. Here, we introduce a protocol for lentiviral vector-mediated transgene expression in murine ES cells. Using lentiviral vectors expressing LacZ, green fluorescent protein, and Cre recombinase, we demonstrate the efficiency and utility of the vectors in ES cell study.
基因导入胚胎干细胞(ES细胞)的主要限制在于:(1)基因传递效率低;(2)整合入宿主基因组后基因表达受到抑制。基于I型人类免疫缺陷病毒(HIV-1)的慢病毒载体已被证明是在ES细胞中实现稳定高效基因表达的优良工具。在此,我们介绍一种在小鼠ES细胞中进行慢病毒载体介导的转基因表达的方案。利用表达LacZ、绿色荧光蛋白和Cre重组酶的慢病毒载体,我们证明了这些载体在ES细胞研究中的效率和实用性。