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Development of second- and third-generation bovine immunodeficiency virus-based gene transfer systems
Meghan Matukonis1, Mengtao Li, Rene P Molina
1Genetic Therapy, a Novartis Company, Gaithersburg, MD 20878, USA.
Human Gene Therapy
|August 7, 2002
Summary
Second-generation bovine immunodeficiency virus (BIV) gene transfer systems were developed. These enhanced BIV vectors efficiently transduce cells and offer a safer alternative to human immunodeficiency virus-based systems.
Area of Science:
- * Molecular Biology
- * Virology
- * Gene Therapy
Background:
- * Lentivirus vectors are effective for gene transfer in various cell types.
- * Early bovine immunodeficiency virus (BIV)-based systems exist but can be improved.
Purpose of the Study:
- * To develop second-generation (three-plasmid) and third-generation (four-plasmid) BIV-based gene transfer systems.
- * To enhance the safety profile of BIV vectors.
Main Methods:
- * Deletion or truncation of accessory genes (vif, vpw, vpy, tmx) and tat from the packaging construct.
- * Separation of Rev expression into a distinct plasmid.
- * Generation of BIV vectors using a minimal transfer vector and VSV-G expression plasmid.
Main Results:
- * Developed functional three- and four-plasmid BIV vector systems.
- * Achieved titers greater than 1 x 10^6 transducing units/mL.
- * Demonstrated efficient transduction of both dividing and nondividing cells.
Conclusions:
- * Accessory genes in BIV are dispensable for functional vector development.
- * Modified BIV vectors possess an improved safety profile.
- * BIV vectors represent a viable alternative to HIV-based gene transfer systems.