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Enhancing and diminishing gene function in human embryonic stem cells

Vallier, L., Rugg-Gunn, P. J., Bouhon, I. A., Andersson, F. K., Sadler, A. J. and Pedersen, R. A. (2004) Enhancing and diminishing gene function in human embryonic stem cells. Stem Cells, 22 (1). pp. 2-11. ISSN 1066-5099

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To link to this item DOI: 10.1634/stemcells.22-1-2


It is widely recognized that gain- and loss-of-function approaches are essential for understanding the functions of specific genes, and such approaches would be particularly valuable in studies involving human embryonic stem (hES) cells. We describe a simple and efficient approach using lipofection to transfect hES cells, which enabled us to generate hES cell lines expressing naturally fluorescent green or red proteins without affecting cell pluripotency. We used these cell lines to establish a means of diminishing gene function using small interfering (si)RNAs, which were effective at knocking down gene expression in hES cells. We then demonstrated that stable expression of siRNA could knock down the expression of endogenous genes. Application of these gain- and loss-of-function approaches should have widespread use, not only in revealing the developmental roles of specific human genes, but also for their utility in modulating differentiation.

Item Type:Article
Divisions:Life Sciences > School of Biological Sciences
ID Code:10631
Uncontrolled Keywords:human embryonic stem cells, fluorescent reporter genes, green, fluorescent protein, red fluorescent protein, transfection, small, interfering RNA, gain-of-function, loss-of-function, stem cell therapy, regenerative medicine, GREEN FLUORESCENT PROTEIN, RNA INTERFERENCE, MAMMALIAN-CELLS, GERMLINE, TRANSMISSION, LENTIVIRAL VECTORS, HUMAN BLASTOCYSTS, TRANSGENIC MICE, EXPRESSION, DIFFERENTIATION, SYSTEM

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