In vivo gene silencing by chemically modified and stable siRNA

摘要

The present invention provides compositions for RNA interference and methods of use thereof. In particular, the invention provides small interfering RNAs (siRNAs) having modification that enhance the stability of the siRNA without a concomitant loss in the ability of the siRNA to participate in RNA interference (RNAi). The invention also provides siRNAs having modification that increase targeting efficiency. Modifications include chemical crosslinking between the two complementary strands of an siRNA and chemical modification of a 3′ terminus of a strand of an siRNA. Preferred modifications are internal modifications, for example, sugar modification, nucleobase modification and/or backbone modifications. Such modifications are also useful, e.g., to improve uptake of the siRNA by a cell. Functional and genomic and proteomic methods are featured. Therapeutic methods are also featured.

主权项

1. A small interfering RNA (siRNA), comprising a sense strand and an antisense strand, wherein the antisense strand is complementary to the sense strand and has a sequence sufficiently complementary to a target mRNA sequence to direct target-specific RNA interference (RNAi), and
wherein the target mRNA sequence comprises a cleavage site for RISC, and wherein the sense strand and the antisense strand comprise uridines, cytidines, adenosines and guanosines with at least one adenosine or guanosine within 2 nucleotides upstream and 9 nucleotides downstream of the cleavage site referencing the antisense strand and at least one adenosine or guanosine that is not within 2 nucleotides upstream and 9 nucleotides downstream of the cleavage site referencing the antisense strand, and wherein the sense strand and the antisense strand are modified by the substitution of each uridine with a 2′-fluoro uridine and each cytidine with a 2′-fluoro cytidine, and wherein the adeno sines and guano sines of the sense strand are 2′-ribonucleotides, and wherein the antisense strand is modified by the substitution of at least one adenosine with 2′-deoxy adenosine or at least one guanosine with 2′-deoxy guanosine within 2 nucleotides upstream and 9 nucleotides downstream of the cleavage site, and wherein the adenosine and guano sine nucleotides of the antisense strand that are not within 2 nucleotides upstream and 9 nucleotides downstream of the cleavage site are unmodified, such that in vivo stability is enhanced as compared to a corresponding unmodified siRNA or such that the target efficiency is enhanced compared to a corresponding unmodified siRNA.