Gene transfer system derived from the caprine arthritis–encephalitis lentivirus

• Published in: Journal of virological methods Vol. 136; no. 1; pp. 177 - 184
• Main Authors: Mselli-Lakhal, Laila, Guiguen, François, Greenland, Timothy, Mornex, Jean-François, Chebloune, Yahia
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 01.09.2006; Amsterdam Elsevier; New York, NY
• Subjects: Arthritis-Encephalitis Virus, Caprine - genetics; Arthritis-Encephalitis Virus, Caprine - growth & development; Biological and medical sciences; Biological Transport; CAEV; Caprine arthritis encephalitis virus; Cell Line; Cytomegalovirus; Cytomegalovirus - genetics; Flow Cytometry; Fundamental and applied biological sciences. Psychology; Gene; Gene Expression; Gene Transfer Techniques; Genes, env; Genes, gag; Genes, pol; Genes, Reporter; Genetic Therapy - methods; Genetic Vectors - genetics; Genetic Vectors - pharmacology; Green Fluorescent Proteins - genetics; Helper Viruses; Human immunodeficiency virus 1; Humans; Lentivirus; Life Sciences; Membrane Glycoproteins - metabolism; Microbiology; Microbiology and Parasitology; Promoter Regions, Genetic; RNA, Viral - metabolism; Techniques used in virology; Transfer; Vector; Vesicular stomatitis virus; Viral Envelope Proteins - metabolism; Virology; Virus Assembly; CAEV; Transfer; Gene; Lentivirus; Vector; Tool; Caprine arthritis encephalitis virus; Microbiology; Retroviridae; Method; Genetic transfer; Virology; Virus; GENE; TRANSFER; TOOL; VECTOR
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2006.05.006

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.