TY - JOUR
T1 - Differences in silencing of mismatched targets by sliced versus diced siRNAs
AU - Sun, Guihua
AU - Wang, Jinghan
AU - Huang, Yasheng
AU - Yuan, Christine Wan Yin
AU - Zhang, Keqiang
AU - Hu, Shuya
AU - Chen, Linling
AU - Lin, Ren Jang
AU - Yen, Yun
AU - Riggs, Arthur D.
N1 - Funding Information:
City of Hope (to A.D.R.); Taiwan Ministry of Health and Welfare Surcharge of Tobacco Products [MOHW105-TDU-B-212-134001 CERC to Y.Y.]. Funding for open access charge: City of Hope (to A.D.R.).
Publisher Copyright:
© The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.
PY - 2018/1/1
Y1 - 2018/1/1
N2 - It has been reported that the two major types of RNA interference triggers, the classical Dicer-generated small RNAs (siRNAs), which function with all members of the Argonaute (Ago) protein family in mammals, and the Ago2-sliced small RNAs (sli-siRNAs), which function solely through Ago2, have similar potency in target cleavage and repression. Here, we show that sli-siRNAs are generally more potent than siRNAs in silencing mismatched targets. This phenomenon is usually more apparent in targets that have mismatched nucleotides in the 3 supplementary region than in targets with mismatches in the seed region. We demonstrate that Ago2 slicer activity is a major factor contributing to the greater silencing efficiency of sli-siRNA against mismatched targets and that participation of non-slicing Agos in silencing mismatched siRNA targets may dilute the slicing ability of Ago2. The difference in length of the mature guide RNA used in sli-RISCs and si-RISCs may also contribute to the observed difference in knockdown efficiency. Our data suggest that a sli-siRNA guide strand is likely to have substantially stronger off-target effects than a guide strand with the same sequence in a classical siRNA and that Dicer and non-slicing Agos may play pivotal roles in controlling siRNA target specificity.
AB - It has been reported that the two major types of RNA interference triggers, the classical Dicer-generated small RNAs (siRNAs), which function with all members of the Argonaute (Ago) protein family in mammals, and the Ago2-sliced small RNAs (sli-siRNAs), which function solely through Ago2, have similar potency in target cleavage and repression. Here, we show that sli-siRNAs are generally more potent than siRNAs in silencing mismatched targets. This phenomenon is usually more apparent in targets that have mismatched nucleotides in the 3 supplementary region than in targets with mismatches in the seed region. We demonstrate that Ago2 slicer activity is a major factor contributing to the greater silencing efficiency of sli-siRNA against mismatched targets and that participation of non-slicing Agos in silencing mismatched siRNA targets may dilute the slicing ability of Ago2. The difference in length of the mature guide RNA used in sli-RISCs and si-RISCs may also contribute to the observed difference in knockdown efficiency. Our data suggest that a sli-siRNA guide strand is likely to have substantially stronger off-target effects than a guide strand with the same sequence in a classical siRNA and that Dicer and non-slicing Agos may play pivotal roles in controlling siRNA target specificity.
UR - http://www.scopus.com/inward/record.url?scp=85055342929&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85055342929&partnerID=8YFLogxK
U2 - 10.1093/nar/gky287
DO - 10.1093/nar/gky287
M3 - Article
AN - SCOPUS:85055342929
SN - 0305-1048
VL - 46
SP - 6806
EP - 6822
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 13
ER -