Compositions and Methods for Non-Targeted Activation of Endogenous Genes
AssigneeABT Holding Company
InventorsJOHN J. HARRINGTON, BRUCE SCHERF, STEPHEN RUNDLETT
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Abstract
The present invention is directed generally to activating gene expression or causing over-expression of a gene by non-homologous or illegitimate recombination, of a regulatory sequence that causes expression of the gene. Thus, by the present invention, 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.
Claims14
1-57. (canceled)
A method for increasing protein expression of an endogenous gene in a eukaryotic cell in vitro, said method comprising introducing a vector into said eukaryotic cell, said vector comprising (i) a first promoter operably linked to a nucleotide sequence encoding a selectable marker, wherein said nucleotide sequence lacks a functional polyadenylation signal, and (ii) a second promoter operably linked to an unpaired splice donor, wherein said vector is non-homologously integrated into the genome of said eukaryotic cell in such a way that a fusion transcript comprising the nucleotide sequence encoding the selectable marker and/or the unpaired splice donor and one or more exons of an endogenous gene is expressed under the control of said first or said second promoter and wherein said unpaired splice donor is spliced to a splice acceptor of said endogenous gene to produce said fusion transcript, and coding sequence in said endogenous gene is translated.
A method for increasing protein expression of an endogenous gene in a eukaryotic cell in vitro, said method comprising introducing a vector into said eukaryotic cell, said vector comprising (i) a first promoter operably linked to a nucleotide sequence encoding a selectable marker, wherein said nucleotide sequence lacks a functional polyadenylation signal, and (ii) a second promoter operably linked to an unpaired splice donor, wherein said vector is non-homologously integrated into the genome of said eukaryotic cell in such a way that a fusion transcript comprising the nucleotide sequence encoding the selectable marker and/or the unpaired splice donor and one or more exons of an endogenous gene is expressed under the control of said first or said second promoter, and coding sequence in said endogenous gene is translated.
Dependencies are inferred from the claim text.