Patent ID: 6933133
Filing Date: 2005-08-23
Classification: A61K,A61P,C07K,C12N

Abstract:
1. A method for obtaining glucose-regulated expression of insulin in vitro in hepatocyte cells, wherein the method comprises delivering a genetic construct for glucose-regulated synthesis of insulin into a host hepatocyte cell in which insulin is not natively produced, wherein the genetic construct comprises (a) a promoter functional in the host hepatocyte cell; (b) a coding sequence for a human proinsulin operably connected to the promoter, wherein the coding sequence encodes a protein which comprises amino acid sequences corresponding to the B-C and C-A junctions of a native human proinsulin; (c) a glucose responsive regulatory module located 5′ of the promoter, wherein the module has at least one glucose inducible regulatory element comprising a pair of CACGTG motifs linked by a five base nucleotide sequence, and wherein the module comprises a sequence selected from the group consisting of SEQ ID No:9 and SEQ ID N:10; and (d) an untranslated region not natively associated with the coding sequence for human proinsulin, wherein the untranslated region is located 5′ of the coding sequence and 3′ of the promoter; wherein glucose-regulated expression of insulin in vitro is achieved.