Generation of High-Titer Defective HSV-1 Amplicon Vectors

Abstract

Strategies for gene delivery into neurons, either to study the molecular biology of brain function or for gene therapy, must utilize vectors that persist stably in postmitotic cells and that can be targeted both spatially and temporally in the nervous system in vivo. Herpes simplex virus type 1 (HSV-1) possesses multiple features that make it an ideal vector for genetic intervention in the nervous system. In particular, it accepts large molecules of exogenous DNA; it infects nondividing cells from a wide range of hosts with high efficiency; it enables the strong expression of foreign genes; it is episomal and thereby does not cause integration effects; its infection of postmitotic cells is stable; and its particles can be concentrated to relatively high titers. Here, we describe the considerations involved in the preparation and use of a herpes simplex virus type 1 (HSV-1) amplicon as a vector for gene transfer into neurons both in vitro and in vivo.