title: Genetic modification of primary chronic lymphocytic leukemia cells with a lentivirus expressing CD38.
abstract: Studies of the role of individual genes in chronic lymphocytic leukemia (CLL) have been hampered by the inability to consistently transfect primary tumor cells. Here, we describe a highly efficient method of genetically modifying primary CLL cells using a VSVG pseudotyped lentiviral vector. We transduced CD38 negative CLL cells with a lentiviral vector encoding CD38 which caused increased surface CD38 expression in all the samples tested (n=17) with no evidence of plasmacytoid differentiation. The mean percentage of positive cells expressing CD38 was 87%+/-8.5% and the mean cell viability 74%+/-17%. This high level of transduction of all the CLL cell samples tested demonstrates the utility of this technique which should prove applicable for the introduction and analysis of other genes in these non-dividing cells.
authors.affiliation: Department of Infection, Immunity and Biochemistry, School of Medicine, Cardiff University, Cardiff, UK.
authors.forename: Laurence
authors.initials: L
authors.lastname: Pearce
authors.forename: Liam
authors.initials: L
authors.lastname: Morgan
authors.forename: Thet Thet
authors.initials: TT
authors.lastname: Lin
authors.forename: Saman
authors.initials: S
authors.lastname: Hewamana
authors.forename: R James
authors.initials: RJ
authors.lastname: Matthews
authors.forename: Silvia
authors.initials: S
authors.lastname: Deaglio
authors.forename: Clare
authors.initials: C
authors.lastname: Rowntree
authors.forename: Christopher
authors.initials: C
authors.lastname: Fegan
authors.forename: Christopher
authors.initials: C
authors.lastname: Pepper
authors.forename: Paul
authors.initials: P
authors.lastname: Brennan
mesh_terms: D051997:ADP-ribosyl Cyclase 1 / Q000235:genetics* / Q000378:metabolism; D049109:Cell Proliferation; D005434:Flow Cytometry; D015973:Gene Expression Regulation, Leukemic; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic
chemical_list: D008562:Membrane Glycoproteins; C498154:CD38 protein, human; D051997:ADP-ribosyl Cyclase 1