Long double-stranded RNA for RNA interference
The present technology relates, in part, to long double-stranded RNA (dsRNA) (e.g., 30 or more base pairs) that inhibits gene expression.
1. A composition comprising long double-stranded RNA (ldRNA) capable of triggering gene silencing via an RNA interference (RNAi) mechanism, wherein
the ldRNA consists of two substantially complementary RNA strands and comprises at least one guide sequence complementary to one or more target mRNAs, and
each of the RNA strands is in the range of 40 to 80 nucleotides and has an overhang on a 3′ end, wherein the ldRNA comprises one or more phosphorothioate linkages.
2. The composition of claim 1 , wherein each of the RNA strands is in the range of 45 to 60 nucleotides.
3. The composition of claim 1 , wherein each of the RNA strands is in the range of 50 to 60 nucleotides.
4. The composition of claim 1 , wherein the ldRNA comprises two or more sequences of at least 8 base pair (bp) having a guide sequence complementary to the same target mRNA.
5. The composition of claim 1 , wherein the ldRNA comprises two or more sequences of at least 8 bp having a guide sequence complementary to different target mRNAs.
6. The composition of claim 1 , wherein an end of the ldRNA comprises a 5′ end of an antisense sequence complementary to the target mRNAs.
7. The composition of claim 1 , wherein the sequences complementary to the target mRNAs are each independently selected from sequences that are 8 to 50 nucleotides in length.
8. The composition of claim 1 , wherein the target mRNAs are viral mRNAs.
9. The composition of claim 1 , wherein the target mRNAs are involved in multiple pathways critical for cancer cell growth and survival, function in the same signal transduction pathway, or function in two or more signal transduction pathways that emanate from a common signal.
10. The composition of claim 1 , wherein the target mRNAs encode an enzyme, a transcription factor, a secreted signaling protein, a signal transduction protein, a kinase or phosphatase, a cellular receptor or an ion channel, or a secreted protein.
11. The composition of claim 1 , wherein the overhang is a di-nucleotide overhang.
12. The composition of claim 1 , wherein the ldRNA causes little or no activation of an antiviral response.
13. A method for inhibiting gene expression comprising administering the composition of claim 1 to a mammalian cell, tissue, or animal.
14. A method of reducing the expression of one or more mRNAs, comprising administering the composition of claim 1 to a mammalian cell, tissue, or animal.
15. A pharmaceutical composition comprising the composition of claim 1 , and a pharmaceutically acceptable carrier.
16. The composition of claim 1 , wherein the ldRNA is encapsulated in liposomes.
17. The composition of claim 1 , wherein the ldRNA is formulated for administration by inhalation, intravenous injection, subcutaneous injection, intradermal injection, intramuscular injection, intraocular injection, intratumoral injection, or other forms of parenteral administration.
18. The composition of claim 1 , wherein the ldRNA is formulated for topical administration.
19. The composition of claim 1 , wherein the ldRNA comprises a lipophilic conjugate selected from cholesterol, tocopherol, cholestene, cholestane, cholestadiene, bile acid, cholic acid, deoxycholic acid, dehydrocholic acid, and a long-chain fatty acid having 10 or more carbon atoms.