Methods and compositions for enhancing the efficacy and specificity of single and double blunt-ended siRNA
First Claim
Patent Images
1. A method of enhancing the ability of a first strand of a RNAi agent to act as a guide strand in mediating RNAi, the RNAi agent derived from an RNA duplex having at least one blunt end, comprising lessening the base pair strength between the 5′
- end of the first strand and the 3′
end of a second strand of the duplex as compared to the base pair strength between the 3′
end of the first strand and the 5′
end of the second strand.
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Abstract
The present invention provides methods of enhancing the efficacy and specificity of RNAi using single or double blunt-ended siRNA. The invention also provides single and double-blunt ended siRNA compositions, vectors, and transgenes containing the same for mediating silencing of a target gene. Therapeutic methods are also featured.
166 Citations
51 Claims
-
1. A method of enhancing the ability of a first strand of a RNAi agent to act as a guide strand in mediating RNAi, the RNAi agent derived from an RNA duplex having at least one blunt end, comprising lessening the base pair strength between the 5′
- end of the first strand and the 3′
end of a second strand of the duplex as compared to the base pair strength between the 3′
end of the first strand and the 5′
end of the second strand. - View Dependent Claims (8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
- end of the first strand and the 3′
-
2. A method of enhancing the efficacy of a siRNA duplex having at least one blunt end, the siRNA duplex comprising a sense and an antisense strand, comprising lessening the base pair strength between the antisense strand 5′
- end (AS 5′
) and the sense strand 3′
end (S 3′
) as compared to the base pair strength between the antisense strand 3′
end (AS 3′
) and the sense strand 5′
end (S '"'"'5), such that efficacy is enhanced.
- end (AS 5′
-
3. A method of enhancing the ability of a first strand of a RNAi agent to act as a guide strand in mediating RNAi, the RNAi agent derived from an RNA duplex having 5′
- and 3′
blunt ends, comprising lessening the base pair strength between the 5′
end of the first strand and the 3′
end of a second strand of the duplex as compared to the base pair strength between the 3′
end of the first strand and the 5′
end of the second strand.
- and 3′
-
4. A method of enhancing the efficacy of a siRNA duplex having 5′
- and 3′
blunt ends, the siRNA duplex comprising a sense and an antisense strand, comprising lessening the base pair strength between the antisense strand 5′
end (AS 5′
) and the sense strand 3′
end (S 3′
) as compared to the base pair strength between the antisense strand 3′
end (AS 3′
) and the sense strand 5′
end (S '"'"'5), such that efficacy is enhanced.
- and 3′
- 5. A method of promoting entry of a desired strand of an siRNA duplex having at least one blunt end into a RISC complex, comprising enhancing the asymmetry of the siRNA duplex, such that entry of the desired strand is promoted.
-
27. A method of decreasing silencing of an inadvertent target mRNA by a dsRNAi agent, the dsRNAi agent comprising a sense strand, an antisense strand, and having at least one blunt end comprising:
-
(a) detecting a significant degree of complementarity between the sense strand and the inadvertent target; and
(b) enhancing the base pair strength between the 5′
end of the sense strand and the 3′
end of the antisense strand relative to the base pair strength between the 3′
end of the sense strand and the 5′
end of the antisense strand;
such that silencing of the inadvertent target mRNA is decreased. - View Dependent Claims (28)
-
-
29. An RNAi agent or siRNA duplex having 5′
- and 3′
blunt ends comprising a sense strand and an antisense strand, wherein the base pair strength between the antisense strand 3′
end (AS 3′
) and the sense strand 5′
end (S 5′
) is less than the base pair strength between the antisense strand 5′
end (AS 5′
) and the sense strand 3′
end (S '"'"'3), such that the antisense strand preferentially guides cleavage of a target mRNA. - View Dependent Claims (30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51)
- and 3′
Specification