RRC ID 4342
著者 Suzuki K, Mitsui K, Aizawa E, Hasegawa K, Kawase E, Yamagishi T, Shimizu Y, Suemori H, Nakatsuji N, Mitani K.
タイトル Highly efficient transient gene expression and gene targeting in primate embryonic stem cells with helper-dependent adenoviral vectors.
ジャーナル Proc Natl Acad Sci U S A
Abstract Human embryonic stem (hES) cells are regarded as a potentially unlimited source of cellular materials for regenerative medicine. For biological studies and clinical applications using primate ES cells, the development of a general strategy to obtain efficient gene delivery and genetic manipulation, especially gene targeting via homologous recombination (HR), would be of paramount importance. However, unlike mouse ES (mES) cells, efficient strategies for transient gene delivery and HR in hES cells have not been established. Here, we report that helper-dependent adenoviral vectors (HDAdVs) were able to transfer genes in hES and cynomolgus monkey (Macaca fasicularis) ES (cES) cells efficiently. Without losing the undifferentiated state of the ES cells, transient gene transfer efficiency was approximately 100%. Using HDAdVs with homology arms, approximately one out of 10 chromosomal integrations of the vector was via HR, whereas the rate was only approximately 1% with other gene delivery methods. Furthermore, in combination with negative selection, approximately 45% of chromosomal integrations of the vector were targeted integrations, indicating that HDAdVs would be a powerful tool for genetic manipulation in hES cells and potentially in other types of human stem cells, such as induced pluripotent stem (iPS) cells.
巻・号 105(37)
ページ 13781-6
公開日 2008-9-16
DOI 10.1073/pnas.0806976105
PII 0806976105
PMID 18768795
PMC PMC2544531
MeSH Adenoviridae / genetics* Animals Cell Line Embryonic Stem Cells / metabolism* Gene Expression / genetics* Gene Targeting / methods* Gene Transfer Techniques* Genetic Vectors / genetics* Humans Hypoxanthine Phosphoribosyltransferase / genetics Hypoxanthine Phosphoribosyltransferase / metabolism Macaca fascicularis / genetics* Mice
IF 9.412
引用数 82
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
リソース情報
ヒトES細胞 KhES-1 KhES-3