MICRORNA-INDUCED ES-LIKE CELLS AND USES THEREOF
The invention relates to isolated nucleic acids comprising mir-302 genes. Also disclosed are expression vectors, host cells, and transgenic animals containing the nucleic acids, and use of the nucleic acids to generate ES-like cells.
1 . An isolated nucleic acid comprising one or more mir (microRNA)-302 genes operably linked to a regulatory sequence, wherein the regulatory sequence controls the expression of the mir-302 genes.
2 . The nucleic acid of claim 1 , wherein the mir-302 genes are selected from the group consisting of the mir-302a, mir-302b, mir-302c, and mir-302d gene.
3 . An isolated nucleic acid comprising a regulatory sequence operably linked to a recombinant sequence encoding a contiguous transcript,
wherein the regulatory sequence controls the transcription of the recombinant sequence,
wherein the recombinant sequence comprises a first gene encoding at least two exons flanking one intron,
wherein the intron comprises one or more mir-302s, and
wherein the intron is spliced out of the contiguous transcript of the recombinant sequence to allow the mir-302s to interact with their targets in a cell.
4 . The nucleic acid Of claim 3 , wherein the mir-302s are selected from the group consisting of mir-302a, mir-302b, mir-302c, and mir-302d.
5 . The nucleic acid of claim 3 , wherein the nucleic acid is transcribed by a type II RNA polymerase.
6 . The nucleic acid of claim 3 , wherein the intron is spliced out of the contiguous transcript of the recombinant sequence by a spliceosome.
7 . The nucleic acid of claim 3 , wherein the first gene is the RGFP (red fluorescent HcRed1 chromoprotein) gene or a fragment thereof.
8 . An expression vector comprising the nucleic acid of claim 1 or 3 .
9 . A host cell comprising the nucleic acid of claim 1 or 3 .
10 . A transgenic animal comprising the nucleic acid of claim 1 or 3 .
11 . A method of generating ES (embryonic stem)-like cells, comprising contacting non-ES-like cells with the nucleic acid of claim 1 or 3 , thereby transforming the non-ES-like cells into ES-like cells.
12 . The method of claim 11 , wherein the non-ES-like cells are cancer cells.
13 . The method of claim 12 , wherein the non-ES-like cells are Colo or PC3 cells.
14 . The method of claim 11 , further comprising inducing the ES-like cells to differentiate into tissue cell types.
15 . The method of claim 14 , wherein the non-ES-like cells are cancer cells.
16 . The method of claim 15 , wherein the non-ES-like cells are Colo or PC3 cells.
17 . The method of claim 16 , wherein the ES-like cells form a teratoma-like primordial tissue structure, fibroblasts, chondrocytes, spermatogonia-like primordial cells, or neuronal cells.