Production of High‐Titer Lentiviral Vectors

Abstract Conventional retroviral vectors are of limited usefulness for neuroscience applications because they are derived from oncoretroviruses such as the Mouse Leukemia Virus (MLV) and, as a consequence, cannot transduce nondividing cells. Most cells in the nervous system proliferate very little, if at all. In contrast to oncoretroviruses, lentiviruses such as the human immunodeficiency virus (HIV) are a sub‐family of retroviruses that can infect both growth‐arrested and dividing cells. Accordingly, lentiviral vectors efficiently transduce targets such as neurons and glial cells, both in tissue culture and in vivo. This unit describes the production of high‐titer lentiviral vectors by transient transfection of 293T cells and concentration of the particles by centrifugation. Guidelines for the titration of vector stocks are also given.