Human Gene Therapy Research Institute

Suming Wang

Charles J Link, Jr

Tatiana Seregina

John P Levy

The present invention describes an efficient retroviral or viral based method that allows easy and quick identification of gene transfer in living, transduced mammalian cells. Retroviral and viral vector producer cells were generated containing a gene for an improved humanized red-shifted, Green Fluorescent Protein (hRGFP) which increases the resulting fluorescence yield after excitation. This humanized, red-shifted GFP (hRGFP) gene was cloned into several vectors and transfected into various packaging cell lines to produce vibrant green fluorescence after excitation with blue light at 450-490 nm. These vectors represent a substantial advance over currently available gene transfer marking systems or wild-type GFP marker systems none of which have been stably transfected into cells.

Vehicles for stable transfer of green fluorescent protein gene and methods of use for same

Disclosed herein is the method for dramatically reducing cytotoxicity of viral vectors such as Herpes simplex viral while retaining gene expression. The method of the invention virtually eliminates the concern of possible recombination during virus propagation and contamination of wild-type virus and virus stock. The invention comprises use of photochemical crosslinking causing differential inactivation of viruses.

Differential inactivation of nucleic acids by chemical modification

A novel HSV mini viral vector is disclosed. The vector comprises HSV and EBV genes which allow it to remain in episomal state, to have very high transfection and infection, and to tolerate up to 140 kb of foreign DNA. Techniques and genetic constructs for producing the vectors, for constructing the vectors and transfection and infection to recipient cells are disclosed.

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Des Moines, IA, United States of America

Suming Wang