TY - JOUR
T1 - Elimination of Off-Target Effect by Chemical Modification of 5'-End of siRNA
T2 - 5′-modified siRNA
AU - Shiohama, Yasuo
AU - Fujita, Ryosuke
AU - Sonokawa, Maika
AU - Hisano, Masaaki
AU - Kotake, Yojiro
AU - Krstic-Demonacos, Marija
AU - Demonacos, Constantinos
AU - Kashiwazaki, Gengo
AU - Fujii, Masayuki
PY - 2022/4/11
Y1 - 2022/4/11
N2 - In this study, the efficiency of RNA interference of small interfering RNAs (siRNAs) bearing 5'-O-methyl-2'-deoxythymidine (X) and 5'-amino-2', 5'-dideoxythymidine (Z) at the 5'-end of the sense strand and the antisense strand of siRNA was investigated in HeLa cells stably expressing enhanced green fluorescent protein. The results indicated that when one strand of siRNA was modified with X or Z and the other was unmodified, the X or Z modification was predominant in the process of strand selection and the unmodified strand was selected as a guide strand. When both strands are modified with X or Z, the modified antisense strand with X or Z will be selected as a guide strand with a certain probability. The resulting mature RNA-induced silencing complex exerted reduced, but still moderate silencing activity remained. These results suggest that the modification of the sense strand with X or Z eliminates the off-target effects caused by the sense strand without affecting the silencing efficiency of the siRNA.
AB - In this study, the efficiency of RNA interference of small interfering RNAs (siRNAs) bearing 5'-O-methyl-2'-deoxythymidine (X) and 5'-amino-2', 5'-dideoxythymidine (Z) at the 5'-end of the sense strand and the antisense strand of siRNA was investigated in HeLa cells stably expressing enhanced green fluorescent protein. The results indicated that when one strand of siRNA was modified with X or Z and the other was unmodified, the X or Z modification was predominant in the process of strand selection and the unmodified strand was selected as a guide strand. When both strands are modified with X or Z, the modified antisense strand with X or Z will be selected as a guide strand with a certain probability. The resulting mature RNA-induced silencing complex exerted reduced, but still moderate silencing activity remained. These results suggest that the modification of the sense strand with X or Z eliminates the off-target effects caused by the sense strand without affecting the silencing efficiency of the siRNA.
KW - 5'-modified siRNA,
KW - RNA, Small Interfering/genetics
KW - HeLa Cells
KW - RNA Interference
KW - RNA, Double-Stranded
KW - RNA-Induced Silencing Complex/metabolism
KW - Thymidine
U2 - 10.1089/nat.2021.0068
DO - 10.1089/nat.2021.0068
M3 - Article
C2 - 35404139
SN - 2159-3337
VL - 32
SP - 438
EP - 447
JO - Nucleic Acid Therapeutics
JF - Nucleic Acid Therapeutics
IS - 5
ER -