Gene transfer system derived from the caprine arthritis-encephalitis lentivirus.
Lentiviruses are attractive candidates for therapeutic vectors, because of their ability to infect non-dividing target cells. Vectors based on HIV-1 efficiently transfer gene expression to a variety of dividing or quiescent cells, but are subject to reservations on safety grounds. Caprine arthritis encephalitis virus (CAEV) is a lentivirus inducing only minor pathology in its natural host and in related species after cross-species transmission. To test the CAEV potential as vector for gene transfer, a cassette expressing the green fluorescent protein (GFP) under control of a CMV promoter was inserted into the CAEV genome, producing the pK2EGFPH vector. When pseudotyped with vesicular stomatitis virus (VSV)-G envelope protein, this vector allowed efficient transfer of GFP expression in human cells (up to 86% of GFP-expressing cells into the TE671 cell line). Three vectors carrying different parts of the viral gag, pol and env genes were then developed, together with a CAEV packaging system. These vectors allowed delimitation of the minimal CAEV sequences necessary for an improvement of vector production compared to the previously described CAEV-based vectors [Mselli-Lakhal et al., 1998. Defect in RNA transport and packaging are responsible for low transduction efficiency of CAEV-based vectors. Arc. Virol. 143, 681-695]. While our previous vectors were produced in a helper/vector system, the present vectors are produced in a helper/free system. However, these vector titers remain lower than those obtained with other lentiviral vectors carrying equivalent packaging sequences. We discuss on possible reasons of such differences and possible improvements.
Mselli-Lakhal
Laila
L
UMR 754 INRA/ENVL/UCBL Retrovirus et pathologie comparée Virologie Cellulaire, Moléculaire et Maladies Emergentes, Université Lyon-1, Bâtiment B; 50, avenue Tony Garnier, 69366 Lyon, Cedex 07, France. lakhal@toulouse.inra.fr
Guiguen
François
F
Greenland
Timothy
T
Mornex
Jean-François
JF
Chebloune
Yahia
Y
D016088:Arthritis-Encephalitis Virus, Caprine / Q000235:genetics* / Q000254:growth & development; D001692:Biological Transport; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics; D005434:Flow Cytometry; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015752:Genes, env; D015750:Genes, gag; D015751:Genes, pol; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics* / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics; D006378:Helper Viruses; D006801:Humans; D008562:Membrane Glycoproteins / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012367:RNA, Viral / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000378:metabolism; D019065:Virus Assembly
C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D012367:RNA, Viral; D014759:Viral Envelope Proteins; D049452:Green Fluorescent Proteins