Improved silencing properties using small internally segmented interfering RNAs

TY - JOUR

T1 - Improved silencing properties using small internally segmented interfering RNAs

AU - Bramsen, Jesper B

AU - Laursen, Maria B

AU - Damgaard, Christian K

AU - Lena, Suzy Willsen

AU - Babu, Bolle Ravindra

AU - Wengel, Jesper

AU - Kjems, Jørgen

PY - 2007

Y1 - 2007

N2 - RNA interference is mediated by small interfering RNAs (siRNAs) that upon incorporation into the RNA-induced silencing complex (RISC) can target complementary mRNA for degradation. Standard siRNA design usually feature a 19-27 base pair contiguous double-stranded region that is believed to be important for RISC incorporation. Here, we describe a novel siRNA design composed of an intact antisense strand complemented with two shorter 10-12 nt sense strands. This three-stranded construct, termed small internally segmented interfering RNA (sisiRNA), is highly functional demonstrating that an intact sense strand is not a prerequisite for RNA interference. Moreover, when using the sisiRNA design only the antisense strand is functional in activated RISC thereby completely eliminating unintended mRNA targeting by the sense strand. Interestingly, the sisiRNA design supports the function of chemically modified antisense strands, which are non-functional within the context of standard siRNA designs. This suggests that the sisiRNA design has a clear potential of improving the pharmacokinetic properties of siRNA in vivo.

AB - RNA interference is mediated by small interfering RNAs (siRNAs) that upon incorporation into the RNA-induced silencing complex (RISC) can target complementary mRNA for degradation. Standard siRNA design usually feature a 19-27 base pair contiguous double-stranded region that is believed to be important for RISC incorporation. Here, we describe a novel siRNA design composed of an intact antisense strand complemented with two shorter 10-12 nt sense strands. This three-stranded construct, termed small internally segmented interfering RNA (sisiRNA), is highly functional demonstrating that an intact sense strand is not a prerequisite for RNA interference. Moreover, when using the sisiRNA design only the antisense strand is functional in activated RISC thereby completely eliminating unintended mRNA targeting by the sense strand. Interestingly, the sisiRNA design supports the function of chemically modified antisense strands, which are non-functional within the context of standard siRNA designs. This suggests that the sisiRNA design has a clear potential of improving the pharmacokinetic properties of siRNA in vivo.

KW - Cell Line, Tumor

KW - Humans

KW - Oligonucleotides

KW - Oligonucleotides, Antisense

KW - RNA Interference

KW - RNA, Guide

KW - RNA, Small Interfering

U2 - 10.1093/nar/gkm548

DO - 10.1093/nar/gkm548

M3 - Journal article

C2 - 17726057

SN - 0305-1048

VL - 35

SP - 5886

EP - 5897

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 17

ER -