| 摘要 |
The present invention is directed generally to 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. In one embodiment of the invention, expression of an endogenous gene is activated or increased following integration into the cell, by non-homologous or illegitimate recombination, of a regulatory sequence that activates expression of the gene. In another embodiment, the expression of the endogenous gene may be further increased by co-integration of one or more amplifiable markers, and selecting for increased copies of the one or more amplifiable markers located on the integrated vector. In another embodiment, the invention is directed to activation of endogenous genes by non-targeted integration of specialized activation vectors, which are provided by the invention, into the genome of a host cell. In one aspect, the invention is directed to a vector that contains a transcriptional regulatory sequence, a positive selectable marker, a negative selectable marker, and an unpaired splice donor site, that functions such that when the vector is integrated into the genome of a cell and splicing occurs between the splice donor on the vector and the splice acceptor in the genome, the positive selectable marker is active and the negative selectable marker is inactive.
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