Selective inhibition of polyglutamine protein expression
The present invention relates to the selective inhibition of protein expression of CAG repeat-related disease proteins such as Huntingtin Disease Protein and Ataxin-3 using double-stranded RNAs and nucleic acid analogs. Chemically-modified RNAs having at least one mismatch as compared to the target CAG repeat sequence are specifically contemplated.
1. A method for selectively inhibiting expression of a disease protein encoded by an mRNA having expanded CAG repeat region comprising contacting a cell that produces said disease protein with a double-stranded RNA of 15-30 bases that targets said expanded CAG repeat region of a disease protein mRNA, and said double-stranded RNA further contains 1, 2, 3, 4 or 5 base mismatches as compared to the targeted expanded CAG repeat, wherein (i) inhibiting is selective for said disease protein expression over expression of a normal form of said disease protein, an mRNA for said normal form which lacks said expanded CAG repeat region, (ii) said double-stranded RNA contains no more than one base mismatch in a seed sequence, and (iii) said double-stranded RNA contains the same number of mismatches to the disease protein mRNA as it does to the normal form of said disease protein mRNA, further comprising wherein one strand of the double-stranded RNA has one of the following sequences:
GCUGCUGAUGCUGCUGCUGTT (SEQ ID NO 1),
GCUGCUGCAGCUGCUGCUGTT (SEQ ID NO 3),
GCUGCUGCUACUGCUGCUGTT (SEQ ID NO 5),
GCUGCUGCUUCUGCUGCUGTT (SEQ ID NO 7),
GCUGCUGCUGAUGCUGCUGTT (SEQ ID NO 9), or
UGCUGCUGCAGCUGCUGCUTT (SEQ ID NO 25).
2. A method for selectively inhibiting expression of a disease protein encoded by an mRNA having expanded CAG repeat region comprising contacting a cell that produces said disease protein with a double-stranded RNA of 15-30 bases that targets said expanded CAG repeat region of a disease protein mRNA, and said double-stranded RNA further contains 1, 2, 3, 4 or 5 base mismatches as compared to the targeted expanded CAG repeat, wherein (i) inhibiting is selective for said disease protein expression over expression of a normal form of said disease protein, an mRNA for said normal form which lacks said expanded CAG repeat region, (ii) said double-stranded RNA contains no more than one base mismatch in a seed sequence, and (iii) said double-stranded RNA contains the same number of mismatches to the disease protein mRNA as it does to the normal form of said disease protein mRNA, further comprising wherein one strand of the double-stranded RNA has one of the following sequences:
GCUGCUGCAACUGCUGCUGTT (SEQ ID NO 11),
GCUGCUGCAAAUGCUGCUGTT (SEQ ID NO 106), or
GCUGCUGAAAAUGCUGCUGTT (SEQ ID NO 108).