Genetic engineering of human embryonic stem cells with lentiviral vectors.
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.
Xiong
Chen
C
Cardiovascular Research Institute, Morehouse School of Medicine, Atlanta, GA 30310, USA.
Tang
Dong-Qi
DQ
Xie
Chang-Qing
CQ
Zhang
Li
L
Xu
Ke-Feng
KF
Thompson
Winston E
WE
Chou
Wayne
W
Gibbons
Gary H
GH
Chang
Lung-Ji
LJ
Yang
Li-Jun
LJ
Chen
Yuqing E
YE
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
D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins