Stem Cell Related Patent Number US6361972

Compositions and methods for non-targeted activation of endogenous genes

Harrington, John J.; Mentor, OH, USA
Sherf, Bruce; Spencer, OH, USA
Rundlett, Stephen; Chagrin Falls, OH, USA

This invention describes methods of activating gene expression or causing over-expression of a gene by recombination methods in situ. The invention also is directed generally to methods for expressing an endogenous gene in a cell at levels higher than those normally found in the cell. Further described are methods of expression of an endogenous gene which is activated or increased following integration into the cell, by non-homologous recombination, of a regulatory sequence that activates expression of the gene. Also included are methods by which the expression of the endogenous gene may be further increased by co-integration of one or more amplifiable markers located on the integrated vector. Also described is the activation of endogenous genes by non-targeted integration of specialized activation vectors, into the genome of a host cell. Methods are further disclosed for the identification, activation, isolation, and/or expression of genes that are undiscoverable by current methods since no target sequence is necessary for integration. Central to the concept is the isolation of nucleic acid molecules (particularly cDNA molecules) encoding a variety of proteins, including transmembrane proteins, and the isolation of cells expressing such transmembrane proteins which maybe heterologous transmembrane proteins. Therapeutic and diagnostic applications are provided for these methods by which endogenous genes, including those associated with human disease and development, may be activated and isolated without prior knowledge of the sequence, structure, function, or expression profile of the genes. This invention represents a continuation of other applications filed by the same assignee and inventors.

Application Number: US2000000481375

Date Filed: 01/10/00

Dated Published: 03/26/02

Assignee: Athersys, Inc., Cleveland, OH, USA