Defective RNA packaging is responsible for low transduction efficiency of CAEV-based vectors

Archives of Virology
L Mselli-LakhalY Chebloune

Abstract

Replication defective retroviral vectors are regularly used for transfer and expression of exogenous genes into dividing cells and in animals. Since lentiviruses are able to infect terminally differentiated and non-dividing cells, their use to produce replication defective vectors may overcome this limitation. We developed two replication-defective lentiviral vectors based on the genome of Caprine Arthritis Encephalitis Virus (CAEV). The first vector (pBNL2) carries the neo and lacZ marker genes. Neo gene is expressed from a genomic RNA and lacZ gene from a subgenomic RNA. The second vector (pCSHL) carries a single fusion gene encoding both phleomycin resistance and beta-galactosidase activity. Replication-competent CAEV was used as helper virus to provide the viral proteins for transcomplementation of these vectors. Our data demonstrated that the genomes of both vectors were packaged into CAEV virions and transduced into goat synovial membrane cells following infection. However, the vector titers remained 3 to 4 logs lower than those of CAEV. Further analysis showed a lack of accumulation of unspliced pBNL2 RNA into the cytoplasm of producer cells resulting in the packaging of pBNL2 sub-genomic RNA only. In contrast, RNA produ...Continue Reading

Citations

Jul 14, 2000·The Journal of Gene Medicine·P MetharomM Q Wei
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Feb 8, 2003·DNA and Cell Biology·Ricardo Quinonez, Richard E Sutton
Nov 4, 2011·Viruses·Stéphanie Durand, Andrea Cimarelli
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Jun 27, 2006·Journal of Virological Methods·Laila Mselli-LakhalYahia Chebloune
Dec 1, 2001·Expert Opinion on Biological Therapy·S PandyaV Planelles
Mar 28, 2007·Journal of Virological Methods·Laila Mselli-LakhalYahia Chebloune
Nov 18, 2017·The Anatomical Record : Advances in Integrative Anatomy and Evolutionary Biology·Chen-Xi ZhengYa-Yun Wang
May 22, 2004·Journal of Biomedical Science·Andrew M L LeverJing Zhao
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