Cotransduction of nondividing cells using lentiviral vectors.
Diseases such as AIDS and cancers may require the introduction of multiple genes into either stem cells or nondividing cells, among others, for therapeutic purposes. Such genes may act at different points of the disease pathway, or may constitute a regulatory loop to bypass or rectify the defective gene or pathway underpinning the disease. Ideally, the therapeutic genes must be transduced together in diverse combinations, and the introduction should occur without constraints. Since lentiviral vectors can transduce both dividing and nondividing cells, they are ideal vehicles to investigate combinatorial gene transfer into diverse cells. In this study, we demonstrate that by using two independent lentiviral vectors, pseudotyped with the protein g of vesicular stomatitis virus, up to four genes can be introduced simultaneously into single dividing and nondividing cells. Up to 45% and 73% of dividing and nondividing cells, respectively, could be transduced with two lentiviral vectors. The efficiency of cotransducing a single cell was the product of the individual transduction efficiencies and suggested the absence of viral interference. Multiple and combinatorial gene transduction using lentiviral vectors may prove useful in gene therapy.
Frimpong
K
K
Department of Pediatrics, University of California, La Jolla 92093-0672, USA.
Spector
S A
SA
D000199:Actins / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D015870:Gene Expression; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D054461:Lim Kinases; D008164:Luminescent Proteins / Q000235:genetics; D008856:Microscopy, Fluorescence; D009474:Neurons; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D013234:Stem Cells; D014162:Transfection / Q000379:methods*; D018980:Williams Syndrome / Q000628:therapy*
D000199:Actins; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D011494:Protein Kinases; C515892:LIMK1 protein, human; D054461:Lim Kinases; D017346:Protein Serine-Threonine Kinases