Specific and stable gene transfer to human embryonic stem cells using pseudotyped lentiviral vectors.
Genetic modification of human embryonic stem cells (hESCs) is an important tool for understanding and influencing their biologic properties. At the present time, lentiviral vectors pseudotyped with the vesicular stomatitis virus G protein (VSV-G) have been most effective for stable gene transfer to hESCs. However, they also efficiently transduce murine embryonic fibroblasts (MEF), used to support the undifferentiated state of many commonly used hESC lines. Transduction of both the MEF as well as hESCs complicates analyses of gene transfer and expression. We made lentiviral vectors pseudotyped with envelope glycoproteins from retroviruses that have been shown to have more restricted transduction ranges and evaluated their specificity. Lentiviral vectors pseudotyped by the envelopes from either the gibbon ape leukemia virus (GALV) or the RD114 feline endogenous virus (RD114) specifically transduced hESCs to similar extents as VSV-G pseudotyped vectors, but did not transduce MEF. In addition, gene modfication by these pseudotyped lentiviral vectors was stably maintained throughout differentiation of hESCs in vitro. These pseudotyped lentiviral vectors may be valuable tools for efficient, specific and stable gene modification of hESCs.
Jang
Jung-Eun
JE
Department of Molecular Microbiology and Immunology, University of Southern California Keck School of Medicine, Los Angeles, CA 90027, USA.
Shaw
Kit
K
Yu
Xiao-Jin
XJ
Petersen
Denise
D
Pepper
Karen
K
Lutzko
Carolyn
C
Kohn
Donald B
DB
D026962:Amino Acid Transport System ASC / Q000235:genetics; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006023:Glycoproteins / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D015778:Minor Histocompatibility Antigens
D026962:Amino Acid Transport System ASC; D006023:Glycoproteins; D015778:Minor Histocompatibility Antigens; C420506:SLC1A5 protein, human; D049452:Green Fluorescent Proteins