Upstream control elements of the proopiomelanocortin gene and their use
View Patent ↗The nucleic acid sequence of the POMC enhancer is disclosed herein. Sequences from the human, rat, rabbit, hamster, mouse, and cow POMC enhancer are disclosed. Hybrid transgenes, comprising a POMC transcriptional control element operably linked to a nucleic acid sequence encoding a marker are also enclosed. In addition, transgenic mice carrying a hybrid transgene including a POMC control element operably linked to a marker are disclosed herein.
1. An isolated proopiomelanocortin enhancer element, comprising a nucleic acid sequence set forth as SEQ ID NO: 15, wherein the isolated proopiomelanocortin enhancer element elevates transcription of a heterologous coding sequence in a host cell.
2. The isolated proopiomelanocortin enhancer element of claim 1 , consisting of a nucleic acid sequence set forth as SEQ ID NO: 15.
3. The isolated proopiomelanocortin enhancer element of claim 1 , operably linked to a promoter and a heterologous nucleic acid sequence, wherein the enhancer element directs expression of the heterologous nucleic acid sequence in a proopiomelanocortin neuron.
4. The isolated proopiomelanocortin enhancer element of claim 1 , comprising a nucleic acid sequence set forth as SEQ ID NO: 10, or SEQ ID NO: 13.
5. The isolated enhancer element of claim 4 , operably linked to a nucleic acid sequence encoding a heterologous polypeptide.
6. The isolated enhancer clement of claim 5 , wherein the nucleic acid sequence encoding a heterologous polypeptide encodes an enzyme, a marker, antibiotic resistance polypeptide or an antigen.
7. The isolated enhancer element of claim 5 , wherein the nucleic acid sequence encoding a polypeptide encodes green fluorescent protein.
8. The isolated enhancer element of claim 1 , operably linked to a heterologous promoter.
9. An expression vector comprising the isolated enhancer element of claim 1 and a promoter.
10. The expression vector of claim 9 , wherein the vector is a viral vector.
11. The expression vector of claim 9 , wherein the vector is a plasmid vector.
12. An isolated host cell transformed with the vector of claim 9 .
13. The isolated host cell of claim 12 , wherein the host cell is a eukaryotic cell.
14. The isolated host cell of claim 12 , wherein the host cell is a prokaryotic cell.
15. An expression cassette comprising the POMC enhancer of claim 1 , and a nucleic acid sequence encoding proopiomelanocortin comprising at least one exon, wherein a polylinker comprising a unique site for a restriction enzyme is inserted into the exon.
16. The isolated proopiomelanocortin enhancer element of claim 1 , wherein the heterologous coding sequence encodes a marker polypeptide.
17. The isolated enhancer element of claim 1 , comprising a nucleic acid sequence set forth as SEQ ID NO: 10.
18. The isolated enhancer element of claim 1 , comprising a nucleic acid sequence forth as SEQ ID NO: 13.
19. An expression vector comprising the isolated enhancer element of claim 16 and a promoter.
20. The vector of claim 19 , wherein the vector is a viral vector.
21. The vector of claim 19 , wherein the vector is a plasmid vector.
22. An isolated host cell transformed with the vector of claim 19 .
23. The isolated host cell of claim 22 , wherein the host cell is a eukaryotic cell.
24. The expression vector of claim 19 , wherein the isolated enhancer element comprises a sequence set forth as SEQ ID NO: 13.
25. An isolated host cell transformed with the vector of claim 19 , wherein the isolated enhancer element comprises a sequence set forth as SEQ ID NO:13.