Asymmetric shorter-duplex siRNA structures trigger efficient gene silencing with reduced nonspecific effects.
Small interfering RNAs (siRNAs) are short, double-stranded RNAs that mediate efficient gene silencing in a sequence-specific manner by utilizing the endogenous RNA interference (RNAi) pathway. The current standard synthetic siRNA structure harbors a 19-base-pair duplex region with 3' overhangs of 2 nucleotides (the so-called 19+2 form). However, the synthetic 19+2 siRNA structure exhibits several sequence-independent, nonspecific effects, which has posed challenges to the development of RNAi therapeutics and specific silencing of genes in research. In this study, we report on the identification of truncated siRNA backbone structures with duplex regions shorter than 19 bp (referred to as asymmetric shorter-duplex siRNAs or asiRNAs) that can efficiently trigger gene silencing in human cell lines. Importantly, this asiRNA structure significantly reduces nonspecific effects triggered by conventional 19+2 siRNA scaffold, such as sense-strand-mediated off-target gene silencing and saturation of the cellular RNAi machinery. Our results suggest that this asiRNA structure is an important alternative to conventional siRNAs for both functional genomics studies and therapeutic applications.
Chang
Chan Il
CI
Department of Chemistry, Pohang University of Science and Technology, Pohang, Korea.
Yoo
Jae Wook
JW
Hong
Sun Woo
SW
Lee
Shi Eun
SE
Kang
Hye Suk
HS
Sun
Xiangao
X
Rogoff
Harry A
HA
Ban
Changill
C
Kim
Soyoun
S
Li
Chiang J
CJ
Lee
Dong-Ki
DK
D001483:Base Sequence; D002460:Cell Line; D005434:Flow Cytometry; D020868:Gene Silencing; D006801:Humans; D034741:RNA, Small Interfering / Q000737:chemistry*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D034741:RNA, Small Interfering