RNA molecules and vectors for gene silencing
The invention relates to short RNA molecules of about 25 nucleotides in length, which comprise antisense RNA complementary to a targeted mRNA and corresponding sense RNA molecules. These short RNA molecules are effective in post-transcriptional gene silencing (PTGS).
1. A composition for introduction into a cell to effect gene silencing, consisting essentially of isolated short antisense RNA molecules (SARMs) and isolated short sense RNA molecules (SSRMs), collectively short RNA molecules (SRMs),
wherein the SSRMs and SARMs are each of a uniform length of 20-24 nucleotides;
wherein said SARMs are complementary to and can base pair with a target RNA, which target RNA is transcribed from a gene that is silenced when said SRMs are present in a cell containing said gene, and said SSRMs correspond to the target RNA;
wherein said gene is endogenous to an organism selected from the group consisting of a plant, a mammal, an avian organism, a reptile, an insect, and a protozoan, or
said target RNA is generated by a pathogen; and
wherein if said SARMs and SSRMs consist of 20 nucleotides, said SARMs and SSRMs are unmodified.
2. The composition of claim 1 wherein said SRMs are of uniform length of 20 nucleotides.
3. An improved composition for introduction into a cell to effect gene silencing, which improved composition comprises isolated short antisense RNA molecules (SARMs) of a uniform length of 20-24 nucleotides;
wherein said SARMs are complementary to and can base pair with a target RNA, and;
the improvement comprising including in said composition isolated short sense RNA molecules (SSRMs) of a uniform length of 20-24 nucleotides, wherein said SSRMs correspond to said target RNA;
wherein said target RNA is transcribed from a gene that is silenced when said SSRMs and SARMs are present in a cell containing said gene, wherein said gene is endogenous to an organism selected from the group consisting of a plant, a mammal, an avian organism, a reptile, an insect, and a protozoan; or said target RNA is generated by a pathogen;
wherein if said SARMs and SSRMs consist of 20 nucleotides, said SARMs and SSRMs are unmodified.
4. The composition of claim 1 wherein said SRMs are of uniform length of 21 nucleotides.
5. The composition of claim 1 wherein said SRMs are of uniform length of 22 nucleotides.
6. The composition of claim 1 wherein said SRMs are of uniform length of 23 nucleotides.
7. The composition of claim 1 wherein said SRMs are of uniform length of 24 nucleotides.
8. The composition of claim 3 wherein the SSRMs and SARMs each consist of 20 nucleotides.
9. The composition of claim 3 wherein the SSRMs and SARMs each consist of 21 nucleotides.
10. The composition of claim 3 wherein the SSRMs and SARMs each consist of 22 nucleotides.
11. The composition of claim 3 wherein the SSRMs and SARMs each consist of 23 nucleotides.
12. The composition of claim 3 wherein the SSRMs and SARMs each consist of 24 nucleotides.
13. A composition for introduction into a cell to effect gene silencing, which composition comprises at least one vector which, when introduced into a cell, produces short antisense RNA molecules (SARMs) and short sense RNA molecules (SSRMs), said SARMs and SSRMs designated, collectively, short RNA molecules (SRMs),
wherein the SSRMs and SARMs are each of a uniform length of 20-24 nucleotides;
wherein said SARMs are complementary to and can base pair with a target RNA, which target RNA is transcribed from a gene that is silenced when said SRMs are present in a cell containing said gene, and said SSRMs correspond to the target RNA;
wherein said gene is endogenous to an organism selected from the group consisting of a plant, a mammal, an avian organism, a reptile, an insect, and a protozoan, or
said target RNA is generated by a pathogen.
14. The composition of claim 13 wherein said SRMs are of uniform length of 20 nucleotides.
15. The composition of claim 13 wherein said SRMs are of uniform length of 21 nucleotides.
16. The composition of claim 13 wherein said SRMs are of uniform length of 22 nucleotides.
17. The composition of claim 13 wherein said SRMs are of uniform length of 23 nucleotides.
18. The composition of claim 13 wherein said SRMs are of uniform length of 24 nucleotides.
19. An improved composition for introduction into a cell to effect gene silencing, which composition comprises at least one vector which, when introduced into a cell, produces short antisense RNA molecules (SARMs) of a uniform length of 20-24 nucleotides;
wherein said SARMs are complementary to and can base pair with a target RNA;
the improvement comprising including in said at least one vector sequences such that short sense RNA molecules (SSRMs) are also produced, wherein said SSRMs are of a uniform length of 20-24 nucleotides and said SSRMs correspond to the target RNA;
which target RNA is transcribed from a gene that is silenced when said SARMs and SSRMs are present in a cell containing said gene,
wherein said gene is endogenous to an organism selected from the group consisting of a plant, a mammal, an avian organism, a reptile, an insect, and a protozoan, or
said target RNA is generated by a pathogen.
20. The composition of claim 19 wherein the SSRMs and SARMs each consist of 20 nucleotides.
21. The composition of claim 19 wherein the SSRMs and SARMs each consist of 21 nucleotides.
22. The composition of claim 19 wherein the SSRMs and SARMs each consist of 22 nucleotides.
23. The composition of claim 19 wherein the SSRMs and SARMs each consist of 23 nucleotides.
24. The composition of claim 19 wherein the SSRMs and SARMs each consist of 24 nucleotides.