DCX mini-promoters
View Patent ↗Isolated polynucleotides comprising a DCX mini-promoters are provided. The mini-promoter may be operably linked to an expressible sequence, e.g. reporter genes, genes encoding a polypeptide of interest, regulatory RNA sequences such as miRNA, siRNA, anti-sense RNA, etc., and the like. In some embodiments a cell comprising a stable integrant of an expression vector is provided, which may be integrated in the genome of the cell. The promoter may also be provided in a vector, for example in combination with an expressible sequence. The polynucleotides find use in a method of expressing a sequence of interest, e.g. for identifying or labeling cells, monitoring or tracking the expression of cells, gene therapy, etc.
1. An isolated polynucleotide comprising a DCX mini-promoter wherein the DCX mini-promoter comprises a DCX regulatory element with substantial similarity to SEQ ID NO: 6, 7 or 8 operably joined to an DCX basal promoter with substantial similarity to SEQ ID NO: 4 or 5 wherein the spacing between the DCX regulatory element and the DCX basal promoter is not more than 500 nucleotides (nt).
2. The polynucleotide of claim 1 comprising a DCX minipromoter with substantially similarity to SEQ ID NO: 1.
3. The polynucleotide of claim 1 comprising a DCX mini-promoter with substantial similarity to SEQ ID NO: 2.
4. The isolated polynucleotide of claim 1 , operably linked to an expressible sequence.
5. A vector comprising the isolated polynucleotide of claim 1 .
6. An isolated cell comprising the vector of claim 5 .
7. The cell of claim 6 , wherein the vector is stably integrated into the genome of the cell.
8. The cell of claim 6 or claim 7 , wherein the cell is a stem cell or a retinal cell.
9. A method of expressing a sequence of interest, the method comprising operably linking the sequence of interest to the polynucleotide of claim 1 ; and introducing into a cell permissive for expression from the DCX mini-promoter.
10. An isolated polynucleotide comprising a DCX mini-promoter wherein the DCX mini-promoter comprises a DCX regulatory element with at least 95% sequence identity to SEQ ID NO: 6, 7 or 8 operably joined to an DCX basal promoter with at least 95% sequence identity to SEQ ID NO: 4 or 5, wherein the spacing between the DCX regulatory element and the DCX basal promoter is not more than 500 nucleotides (nt).
11. The polynucleotide of claim 10 , wherein the DCX mini-promoter has at least 95% sequence identity to SEQ ID NO: 1.
12. The polynucleotide of claim 10 wherein the DCX mini-promoter has at least 95% sequence identity to SEQ ID NO: 2.
13. The isolated polynucleotide of claim 10 , operably linked to an expressible sequence.
14. A vector comprising the isolated polynucleotide of claim 10 .
15. An isolated cell comprising the vector of claim 14 .
16. The cell of claim 15 , wherein the vector is stably integrated into the genome of the cell.
17. The cell of claim 15 , wherein the cell is a stem cell or a retinal cell.
18. A method of expressing a sequence of interest, the method comprising operably linking the sequence of interest to the polynucleotide of claim 10 ; and introducing into a cell permissive for expression from the DCX mini-promoter.