4.0 Article

Targeted Gene Modification in Mouse ES Cells Using Integrase-Defective Lentiviral Vectors

期刊

GENESIS
卷 47, 期 4, 页码 217-223

出版社

WILEY
DOI: 10.1002/dvg.20469

关键词

lentivirus; gene targeting; knockout; gene therapy; homologous recombination

资金

  1. Astellas Foundation for Research on Metabolic Disorders
  2. Nakajima Foundation
  3. MEXT of JAPAN
  4. Grants-in-Aid for Scientific Research [20670006] Funding Source: KAKEN

向作者/读者索取更多资源

Lentiviral vectors efficiently integrate into the host genome of both dividing and nondividing cells, and so they have been used for stable transgene expression in biological and biomedical studies. However, recent studies have highlighted the risk of insertional mutagenesis and subsequent oncogenesis. Here, we used an integrase-defective lentiviral (IDLV) vector to decrease the chance of random integration and examined the feasibility of lentiviral vector-mediated gene targeting into murine embryonic stem (ES) cells. After transduction with wild-type lentiviral vectors, none of the 512 G418 resistant clones were found to be homologous recombinant clones. Although the transduction efficiency was lower with the IDLV vectors (5.9% of wild-type), successful homologous recombination was observed in nine out of the 941 G418 resistant clones (0.83 +/- 1.32%). Pluripotency of the homologous recombinant ES cells was confirmed by the production of chimeric mice and subsequent germ line transmission. Because lentiviral vectors can efficiently transduce a variety of stem cell types, our strategy has potential relevance for secure gene-manipulation in therapeutic applications. genesis 47:217-223, 2009. (C) 2009 Wiley-Liss, Inc.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.0
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据