Elimination of Off-Target Effect by Chemical Modification of 5'-End of Small Interfering RNA.

IF 4 2区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Nucleic acid therapeutics Pub Date : 2022-04-11 DOI:10.1089/nat.2021.0068

Yasuo Shiohama, Ryosuke Fujita, Maika Sonokawa, Masaaki Hisano, Y. Kotake, M. Krstic-Demonacos, C. Demonacos, Gengo Kashiwazaki, T. Kitayama, M. Fujii

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引用次数: 2

Abstract

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.

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小干扰RNA 5′端化学修饰消除脱靶效应。

本研究在HeLa细胞中稳定表达增强的绿色荧光蛋白，研究了siRNA的5'- o-甲基-2'-脱氧胸腺嘧啶(X)和5'-氨基-2'，5'-二脱氧胸腺嘧啶(Z)的小干扰RNA (siRNA)在siRNA的5'端和反义链上的RNA干扰效率。结果表明，当一条siRNA被X或Z修饰而另一条未被修饰时，X或Z修饰在链选择过程中占优势，未修饰的链被选择作为导链。当两条链都被X或Z修饰时，带有X或Z修饰的反义链将以一定的概率被选择为导链。由此产生的成熟rna诱导的沉默复合体的沉默活性降低，但仍保持适度的沉默活性。这些结果表明，用X或Z修饰感链可以消除由感链引起的脱靶效应，而不会影响siRNA的沉默效率。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Nucleic acid therapeutics BIOCHEMISTRY & MOLECULAR BIOLOGY-CHEMISTRY, MEDICINAL

CiteScore

7.60

自引率

7.50%

发文量

审稿时长

>12 weeks

期刊介绍： Nucleic Acid Therapeutics is the leading journal in its field focusing on cutting-edge basic research, therapeutic applications, and drug development using nucleic acids or related compounds to alter gene expression. The Journal examines many new approaches for using nucleic acids as therapeutic agents or in modifying nucleic acids for therapeutic purposes including: oligonucleotides, gene modification, aptamers, RNA nanoparticles, and ribozymes.