Use of RNAi technology to inhibit ASIC3
In vitro studies using cells transfected with acid-sensing ion channel 3 (ASIC3) or acid-sensing ion channel 1 (ASIC1) cDNA, demonstrated that the miRNAs against mouse ASIC3 (miR844 and miR847) selectively inhibit mouse ASIC3, but not ASIC1 as detected by protein expression and responses to pH. When the RNAi agents, miR844 or miR847, were used in vivo, delivered into the muscle of mice using a replication-defective herpes simplex viral (HSV-1) vector, primary and secondary hyperalgesia were reduced after carrageenan-induced muscle inflammation. Accordingly, the present invention provides RNAi agents that target ASIC3, methods of preparing such RNAi agents, and methods of using them to modulate in a cell the level of ASIC3 or activity of an ASIC including at least one ASIC3. Modulation of ASIC3 activity or levels can be used for different purposes such as treating pain associated with the expression of ASIC3 and the like.
1. An isolated oligonucleotide comprising a polynucleotide sequence comprising about 12 to 25 nucleotides in length sufficiently complementary to a microRNA target sequence of ASIC3, wherein the oligonucleotide is capable of decreasing ASIC3 expression or activity, wherein the isolated oligonucleotide comprises a polynucleotide sequence obtained from SEQ ID NOS:1-4, 6-16 or 21 or its complement.
2. An isolated oligonucleotide comprising a polynucleotide sequence comprising about 12 to 25 nucleotides in length sufficiently complementary to a microRNA target sequence of ASIC3, wherein the oligonucleotide is capable of decreasing ASIC3 expression or activity, wherein microRNA target sequence of ASIC3 is the sequence of SEQ ID NO:5, 17, 18, 19, or 20 or a complement thereof.
3. An isolated single stranded microRNA molecule, said molecule comprising the sequence of bases identified in SEQ ID NOS:1, 2, 4, 5, 8, 11, 14, 17-27 or the complement thereof wherein the molecule is capable of decreasing ASIC3 expression or activity.