title: Lentiviral manipulation of gene expression in human adult and embryonic stem cells.
abstract: Human stem cells could revolutionize the field of medicine by providing a diverse range of cell types for tissue replacement therapies and drug discovery. To achieve this goal, genetic tools need to be optimized and developed for controlling and manipulating stem cells ex vivo. Here we describe a lentiviral delivery system capable of high infection rates in human mesenchymal and embryonic stem cells. The lentiviral backbone was modified to express mono- and bi-cistronic transgenes and was also used to deliver short hairpin ribonucleic acid for specific silencing of gene expression in human stem cells. We show that lentiviral transduction can be used to alter gene expression without altering the genes' ability to differentiate in vitro. These vectors will enable rapid analysis of gene function in stem cells and permit the generation of knock-in / knock-out models of human disease in the rapidly developing field of gene therapy.
authors.affiliation: Wolfson Institute of Biomedical Research, University College, London, United Kingdom.
authors.forename: Mark O
authors.initials: MO
authors.lastname: Clements
authors.forename: Andrew
authors.initials: A
authors.lastname: Godfrey
authors.forename: Joanne
authors.initials: J
authors.lastname: Crossley
authors.forename: Sam J
authors.initials: SJ
authors.lastname: Wilson
authors.forename: Yasu
authors.initials: Y
authors.lastname: Takeuchi
authors.forename: Chris
authors.initials: C
authors.lastname: Boshoff
mesh_terms: D000328:Adult; D002460:Cell Line; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D020868:Gene Silencing; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics
chemical_list: D034741:RNA, Small Interfering