Improved silencing properties using small internally segmented interfering RNAs.
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.
Bramsen
Jesper B
JB
Department of Molecular Biology, University of Aarhus, DK-8000, Aarhus C, Denmark.
Laursen
Maria B
MB
Damgaard
Christian K
CK
Lena
Suzy W
SW
Babu
B Ravindra
BR
Wengel
Jesper
J
Kjems
Jørgen
J
D045744:Cell Line, Tumor; D006801:Humans; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense / Q000737:chemistry; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000097:blood / Q000737:chemistry* / Q000378:metabolism; D058727:RNA, Small Untranslated
D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D034741:RNA, Small Interfering; C477371:locked nucleic acid; D058727:RNA, Small Untranslated