TY - JOUR
T1 - Designing Ago2-specific siRNA/shRNA to avoid competition with endogenous miRNAs
AU - Ma, Hongming
AU - Zhang, Junli
AU - Wu, Haoquan
PY - 2014/7/1
Y1 - 2014/7/1
N2 - Relatively large amounts of transfected siRNA can compete for Ago proteins and thus compromise endogenous miRNA function, potentially leading to toxicities. Here, we show that shRNA can also perturb endogenous miRNA function similarly. More importantly, we also show that the problem can be solved by designing shRNAs in the context of pre-miR-451 structure with completely complementary stem, which significantly improves the Ago2 specificity. This shRNA was shown to be Ago2-specific, and maintain target-silencing ability while avoiding competition with endogenous miRNAs by not competing for Agos 1, 3, and 4. We conclude that modified pre-miR-451 structure provides a general platform to design shRNAs that significantly reduce perturbation of miRNA function.
AB - Relatively large amounts of transfected siRNA can compete for Ago proteins and thus compromise endogenous miRNA function, potentially leading to toxicities. Here, we show that shRNA can also perturb endogenous miRNA function similarly. More importantly, we also show that the problem can be solved by designing shRNAs in the context of pre-miR-451 structure with completely complementary stem, which significantly improves the Ago2 specificity. This shRNA was shown to be Ago2-specific, and maintain target-silencing ability while avoiding competition with endogenous miRNAs by not competing for Agos 1, 3, and 4. We conclude that modified pre-miR-451 structure provides a general platform to design shRNAs that significantly reduce perturbation of miRNA function.
KW - Ago2
KW - Dicer
KW - MiR-451
KW - ShRNA
UR - http://www.scopus.com/inward/record.url?scp=84903791789&partnerID=8YFLogxK
U2 - 10.1038/mtna.2014.27
DO - 10.1038/mtna.2014.27
M3 - Article
C2 - 25025466
AN - SCOPUS:84903791789
SN - 2162-2531
VL - 3
SP - e176
JO - Molecular Therapy - Nucleic Acids
JF - Molecular Therapy - Nucleic Acids
M1 - e176
ER -