EXPRESSION OF ENDOGENOUS GENES BY NON-HOMOLOGOUS RECOMBINATION OF A VECTOR CONSTRUCT WITH CELLULAR DNA

摘要

The present invention relates generally to activating gene expression orcausing over-expression of a gene by recombination methods in situ. Theinvention also relates generally to methods for expressing an endogenous genein a cell at levels higher than those normally found in the cell. In oneembodiment of the invention, expression of an endogenous gene is activated orincreased following integration into the cell, by non-homologous orillegitimate recombination, of a regulatory sequence that activates expressionof the gene. In another embodiment, the expression of the endogenous gene maybe further increased by co-integration of one or more amplifiable markers, andselecting for increased copies of the one or more amplifiable markers locatedon the integrated vector. The invention also provides methods for theidentification and expression of genes undiscoverable by current methods sinceno target sequence is necessary for integration. The invention also providesmethods for isolation of nucleic acid molecules (particularly cDNA molecules)encoding transmembrane proteins, and for isolation of cells expressing suchtransmembrane proteins which may be heterologous transmembrane proteins. Theinvention also relates to isolated genes, gene products, nucleic acidmolecules, and compositions comprising such genes, gene products and nucleicacid molecules, that may be used in a variety of therapeutic and diagnosticapplications. Thus, by the present invention, endogenous genes, includingthose associated with human disease and development, may be activated andisolated without prior knowledge of the sequence, structure, function, orexpression profile of the genes.