Integrase-mediated nonviral gene transfection with enhanced integration efficiency.
Retroviruses efficiently integrate their genome into the host chromosome. Two elements of the retrovirus genome are needed for the integration: long terminal repeats (LTRs) and integrase protein. We attempted to incorporate the retrovirus integration machinery in lipid vesicle-mediated gene transfection with the aim of achieving efficient stable transfection in a nonviral gene transfection system. A DNA fragment, in which a neomycin-resistant gene was flanked between partial LTR sequences derived from the Rous sarcoma virus (RSV), was constructed. This DNA fragment was transfected together with purified recombinant RSV integrase or integrase expression vectors by means of lipid vesicle-mediated gene transfection. The integrase-mediated transfection enhanced the stable transfection efficiency. The length and the end structure of the LTR sequences were important in achieving high efficiency. Under optimal conditions, the stable transfection efficiency showed a 16-fold improvement over that without integrase.
Mizuarai
S
S
Department of Biotechnology, Graduate School of Engineering, Nagoya University, Nagoya 464-8603, Japan.
Kamihira
M
M
Nishijima
K
K
Iijima
S
S