Development of Second- and Third-Generation Bovine Immunodeficiency Virus-Based Gene Transfer Systems

Abstract

Lentivirus-based gene transfer systems have demonstrated their utility in mediating gene transfer to dividing and nondividing cells both in vitro and in vivo. An early-generation gene transfer system developed from bovine immunodeficiency virus (BIV) has been described (Berkowitz et al., J. Virol. 2001;75:3371-3382). In this paper, we describe the development of second-generation (three-plasmid) and third-generation (four-plasmid) BIV-based systems. All accessory genes (vif, vpw, vpy, and tmx) and the regulatory gene tat were deleted or largely truncated from the packaging construct. Furthermore, we split the packaging function into two constructs by expressing Rev in a separate plasmid. Together with our minimal BIV transfer vector construct and a vesicular stomatitis virus G glycoprotein-expressing plasmid, the BIV vectors were generated. The vectors produced by the three- and four-plasmid systems had titers greater than 1 x 10(6) transducing units per milliliter and were fully functional as indicated by their ability to efficiently transduce both dividing and nondividing cells. These results suggest that the accessory genes vif, vpw, vpy, and tmx are dispensable for functional BIV vector development. The modifications made to the packaging constructs improve the safety profile of the vector system. Finally, BIV vectors provide an alternative to human immunodeficiency virus-based gene transfer systems.

FIV: from lentivirus to lentivector

“…The potential to exploit this property has been the principal incentive for developing viral vectors from these pathogenic complex retroviruses. After HIV-1 vectors were described, replication-defective systems were engineered from molecular clones of non-primate and other primate lentiviruses [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19]. In addition to their use in pre-clinical gene therapy research, lentiviral vectors have been important tools for basic science [20][21][22][23][24][25][26][27][28].…”

Production of Retroviral and Lentiviral Gene Therapy Vectors: Challenges in the Manufacturing of Lipid Enveloped Virus

“…In addition to HIV-derived, other lentiviral vectors have been developed and reported to retain identical features to those of HIV's based, including the ability to transduce nondividing cells, high titers production, and the possibility to be pseudotyped with different envelope glycoproteins. These include lentiviral vectors based on SIV (simian immunodeficiency virus) (Pandya et al 2001;Schnell et al 2000), BIV (bovine immunodeficiency virus) (Matukonis et al 2002;Molina et al 2004), FIV (feline immunodeficiency virus) (Poeschla et al 1998;Saenz and Poeschla 2004) and EAIV (equine infectious anaemia virus) (Balaggan et al 2006;Mitrophanous et al 1999;Stewart et al 2009). Most of non-HIV derived lentiviral vectors have been reported to be tat and sometimes rev independent, thus falling in the 3 rd or 4 th generation of packaging systems.…”

New Vectors for Stable and Safe Gene Modification

“…Bovine immunodeficiency virus based vectors. Another alternative to HIV-1 based vectors are the LVs based on bovine immunodeficiency virus (BIV) (Berkowitz et al, 2001;Matukonis et al, 2002). BIV-based vectors also transduced a wide panel of gene therapy primary targets including unstimulated human HSCs.…”