METHODS OF INDUCING INSULIN PRODUCTION
The present invention provides a method of inducing insulin production in a cell by up-regulating a target gene involved in insulin production in said cell using an saRNA (short activating ribonucleic acid) which specifically targets a target antisense RNA transcript present in said cell.
1 . An saRNA which is capable of inducing insulin production in a cell by up-regulating a target gene involved in insulin production in said cell by specifically targeting a target RNA transcript present in said cell, wherein the target gene is selected from the group consisting of insulin, PDX1, Rfx6, GLP1 receptor, FOXA2, HNF1A, Hlxb9, Hnf6, Ptf1a, Neuro D, betacellulin, ABCC8, glucokinase (GCK), GLUT2, and Nkx6-1, and wherein
(i) said target RNA transcript is complementary to a sequence located on the coding strand of the target gene between 500 nucleotides upstream and 500 nucleotides downstream of the transcription start site of the target gene; and
(ii) said saRNA comprises a strand of up to 30 nucleotides in length comprising a sequence of at least 13 nucleotides which has at least 95% complementarity to a region of the target transcript.
2 . The saRNA of claim 1 , wherein said target RNA transcript overlaps the transcription start site and/or a promoter region of the target gene.
3 . The saRNA of claim 1 , wherein said saRNA is a single-stranded RNA.
4 . The saRNA of claim 1 , wherein said saRNA is a double-stranded RNA.
5 . The saRNA of claim 4 , wherein each strand of the double-stranded RNA has 19 to 25 base pairs in length with 1-5 unpaired bases on each 3′ end of the strand forming a 3′ overhang.
6 . The saRNA of claim 5 , wherein said 3′ overhand is formed of 2 uracils.
7 . The saRNA of claim 1 , wherein said saRNA possesses none of the motifs aaaa, cccc, gggg, and uuuu and/or has a GC-percentage of at least 20% and no more than 75%.
8 . The saRNA of claim 1 , wherein the GC-percentage of said saRNA is between 20% and 55%.
9 . The saRNA of claim 1 , comprising at least one modification.
10 . The saRNA of claim 1 , comprising a sequence selected from anyone of SEQ ID Nos. 1-4, 7-10, or 15-44.
11 . The saRNA of claim 1 , wherein the cell is induced to produce insulin in a glucose-responsive manner.
12 . The saRNA of claim 1 , wherein the cell is induced to secrete insulin.
13 . The saRNA of claim 1 , wherein the cell is an adult or embryonic stem cell, a pluripotent cell, a multipotent cell, or a somatic cell.
14 . The saRNA of claim 1 , wherein the cell is a liver cell or a pancreatic cell.
15 . A pharmaceutical composition comprising one or more saRNAs according to claim 1 .
16 . A method of inducing insulin production in a cell by up-regulating a target gene involved in insulin production in said cell, wherein the target gene is selected from the group consisting of insulin, PDX1, Rfx6, GLP1 receptor, FOXA2, HNF1A, Hlxb9, Hnf6, Ptf1a, Neuro D, betacellulin, ABCC8, glucokinase (GCK), GLUT2, and Nkx6-1, comprising administering the saRNA of claim 1 .