Adenovirus-transfected primary cells and methods of pathway mapping
AssigneesSanofi-Aventis U.S. LLC, Aventis Pharmaceuticals Inc.
InventorsChang Hahn, Li Li
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Abstract
A primary cell culture is transfected with a vector comprising a reporter gene operatively linked to a cis-element. Expression of a candidate regulatory protein is induced in the cell culture and its effects on the cis-element are assayed.
Claims14
A method of determining whether a stimulus is capable of activating a candidate cis-acting regulatory element in an immunocyte, wherein said cis-acting regulatory element is regulated by at least one transcription factor or enhancer, and wherein said stimulus is known to modulate expression of a signaling pathway, said method comprising the steps of:
- (a) transfecting said immunocyte with a recombinant adenovirus, said recombinant adenovirus comprising a reporter gene operatively linked to said candidate cis-acting regulatory element;
- (b) measuring a base level of reporter gene activity;
- (c) applying said stimulus to said immunocyte;
- and
- (d) measuring reporter gene activity in response to said stimulus.
A method of inhibiting expression of a signaling pathway in an immunocyte comprising the steps of:
- (a) transfecting said immunocyte with a recombinant adenovirus, wherein said recombinant adenovirus comprises a suppressor gene operatively linked to a cis-acting regulatory element, wherein said cis-acting regulatory element belongs to said signaling pathway;
- and
- (b) inducing expression of said suppressor gene.
Dependencies are inferred from the claim text.