title: Genetic engineering of human embryonic stem cells with lentiviral vectors.
abstract: Human embryonic stem (hES) cells present a valuable source of cells with a vast therapeutic potential. However, the low efficiency of directed differentiation of hES cells remains a major obstacle in their uses for regenerative medicine. While differentiation may be controlled by the genetic manipulation, effective and efficient gene transfer into hES cells has been an elusive goal. Here, we show stable and efficient genetic manipulations of hES cells using lentiviral vectors. This method resulted in the establishment of stable gene expression without loss of pluripotency in hES cells. In addition, lentiviral vectors were effective in conveying the expression of an U6 promoter-driven small interfering RNA (siRNA), which was effective in silencing its specific target. Taken together, our results suggest that lentiviral gene delivery holds great promise for hES cell research and application.
authors.affiliation: Cardiovascular Research Institute, Morehouse School of Medicine, Atlanta, GA 30310, USA.
authors.forename: Chen
authors.initials: C
authors.lastname: Xiong
authors.forename: Dong-Qi
authors.initials: DQ
authors.lastname: Tang
authors.forename: Chang-Qing
authors.initials: CQ
authors.lastname: Xie
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.forename: Ke-Feng
authors.initials: KF
authors.lastname: Xu
authors.forename: Winston E
authors.initials: WE
authors.lastname: Thompson
authors.forename: Wayne
authors.initials: W
authors.lastname: Chou
authors.forename: Gary H
authors.initials: GH
authors.lastname: Gibbons
authors.forename: Lung-Ji
authors.initials: LJ
authors.lastname: Chang
authors.forename: Li-Jun
authors.initials: LJ
authors.lastname: Yang
authors.forename: Yuqing E
authors.initials: YE
authors.lastname: Chen
mesh_terms: D002454:Cell Differentiation; D002469:Cell Separation; D004622:Embryo, Mammalian / Q000166:cytology*; D005434:Flow Cytometry; D005818:Genetic Engineering / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D016086:Lentivirus / Q000235:genetics*; D008856:Microscopy, Fluorescence; D008957:Models, Genetic; D032383:Myocytes, Cardiac / Q000166:cytology; D034741:RNA, Small Interfering / Q000378:metabolism; D012038:Regeneration; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology*; D013724:Teratoma / Q000378:metabolism / Q000473:pathology; D014162:Transfection; D019076:Transgenes
chemical_list: D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins