Multimerized enhancer domains for cell-specific expression
AssigneeThe McLean Hospital Corporation
InventorsKwang-Soo Kim, Ole Isacson, Dong-Youn Hwang
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Abstract
The invention features an enhancer cassette and methods of its use. The enhancer cassette has the formula [X−Y]n, wherein each X is independently a cell type-specific enhancer element; Y is absent or is a mono or polynucleotide that has between one and thirty nucleotides; and n is an integer between five and fifty, inclusive.
Claims16
An enhancer cassette having the formula [X−Y]n, wherein each X is independently a cell type-specific enhancer;
- Y is absent or is a mono or polynucleotide that has between one and thirty nucleotides;
- and n is an integer between five and fifty, inclusive.
An expression vector comprising
- (i) an enhancer cassette having the formula [X−Y]n, wherein each X is, independently, is a cell-specific enhancer derived from a TH gene;
- Y is absent or is a mono or polynucleotide that has between one and thirty nucleotides;
- and n is an integer between five and fifty, inclusive;
- (ii) an RNA polymerase binding site;
- (iii) a transcription initiation site;
- and
- (iv) a polynucleotide to be expressed,
- wherein said enhancer cassette, RNA polymerase, transcription initiation site, and polynucleotide are spatially arranged in said vector such that said polynucleotide is capable of being expressed when said vector is in a dopaminergic cell.
A method of expressing a polynucleotide in a dopaminergic cell, comprising introducing into said dopaminergic cell an expression vector comprising:
- (i) an enhancer cassette having the formula [X−Y]n operably linked to said polynucleotide, wherein each X is independently a dopaminergic cell-specific enhancer;
- Y is absent or is a mono or polynucleotide that has between one and thirty nucleotides;
- and n is an integer between five and fifty, inclusive
- (ii) an RNA polymerase binding site;
- (iii) a transcription initiation site;
- and
- (iv) a polynucleotide encoding a polypeptide,
- wherein said enhancer cassette, RNA polymerase, transcription initiation site, and polynucleotide are spatially arranged in said vector such that said polynucleotide is expressed when said vector is in a dopaminergic cell following instriduction of said expression vector into said cell.
A substantially pure polynucleotide consisting essentially of SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, or SEQ ID NO:
- 4.
An enhancer cassette having the formula [X−Y]n, wherein each repeated X subunit, independently, is a polynucleotide selected from SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, and SEQ ID NO:
- 4, each repeated Y subunit, independently, is a polynucleotide containing between zero and thirty nucleotides;
- and n is an integer between five and fifty, inclusive.
Dependencies are inferred from the claim text.