siRNA sequence-independent modification formats for reducing off-target phenotypic effects in RNAi, and stabilized forms thereof
AssigneeApplied Biosystems, LLC
InventorsNitin Puri, Irudaya Charles, Susan Magdaleno, Alexander Vlassov, Chris Burnett
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Abstract
Modification formats having modified nucleotides are provided for siRNA. Short interfering RNA having modification formats and modified nucleotides provided herein reduce off-target effects in RNA interference of endogenous genes. Further modification formatted siRNAs are demonstrated to be stabilized to nuclease-rich environments. Unexpectedly, increasing or maintaining strand bias, while necessary to maintain potency for endogenous RNA interference, is not sufficient for reducing off-target effects in cell biology assays.
Claims15
A chemically synthesized short interfering RNA comprising:
- a chemically synthesized passenger oligonucleotide having a length of 15 to 30 nucleotides and comprising:
- sequence-independent modification format (1):
- (1) 5′ Np-m-m-Nx-m-m-Nq-nr 3′, wherein p is 0;
- x is 10;
- r is 0, 1, or 2;
- and q is an integer representing the length of the passenger strand minus (p+x+r+4);
- wherein
- each m is independently a bicyclo nucleotide or a tricyclo nucleotide;
- each n is independently a deoxynucleotide, modified nucleotide, or ribonucleotide;
- when m is a bicyclo nucleotide, each N is independently a nucleotide other than a bicyclo nucleotide, and
- when m is a tricyclo nucleotide, each N is independently a nucleotide other than a tricyclo nucleotide
- and
- a guide oligonucleotide having 15 to 30 nucleotides including a region of continuous complementarity to the passenger oligonucleotide of at least 12 nucleotides,
- the guide oligonucleotide further having complementarity to at least a portion of a target mRNA.
Dependencies are inferred from the claim text.