Investigation of Strand-Selective Interaction of SNA-Modified siRNA with AGO2-MID
Small interfering RNA (siRNA) has been recognized as a powerful gene-silencing tool. For therapeutic application, chemical modification is often required to improve the properties of siRNA, including its nuclease resistance, activity, off-target effects, and tissue distribution. Careful siRNA guide...
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MDPI AG
2020-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/15/5218 |
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| author | Yukiko Kamiya Yuuki Takeyama Tomonari Mizuno Fuminori Satoh Hiroyuki Asanuma |
| author_facet | Yukiko Kamiya Yuuki Takeyama Tomonari Mizuno Fuminori Satoh Hiroyuki Asanuma |
| author_sort | Yukiko Kamiya |
| collection | DOAJ |
| description | Small interfering RNA (siRNA) has been recognized as a powerful gene-silencing tool. For therapeutic application, chemical modification is often required to improve the properties of siRNA, including its nuclease resistance, activity, off-target effects, and tissue distribution. Careful siRNA guide strand selection in the RNA-induced silencing complex (RISC) is important to increase the RNA interference (RNAi) activity as well as to reduce off-target effects. The passenger strand-mediated off-target activity was previously reduced and on-target activity was enhanced by substitution with acyclic artificial nucleic acid, namely serinol nucleic acid (SNA). In the present study, the reduction of off-target activity caused by the passenger strand was investigated by modifying siRNAs with SNA. The interactions of SNA-substituted mononucleotides, dinucleotides, and (2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO)-labeled double-stranded RNA (dsRNA) with the MID domain of the Argonaute 2 (AGO2) protein, which plays a pivotal role in strand selection by accommodation of the 5’-terminus of siRNA, were comprehensively analyzed. The obtained nuclear magnetic resonance (NMR) data revealed that AGO2-MID selectively bound to the guide strand of siRNA due to the inhibitory effect of the SNA backbone located at the 5’ end of the passenger strand. |
| first_indexed | 2024-03-10T18:16:21Z |
| format | Article |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T18:16:21Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-cc0fc757bbf94276a617ab3a5669f7952023-11-20T07:42:01ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-07-012115521810.3390/ijms21155218Investigation of Strand-Selective Interaction of SNA-Modified siRNA with AGO2-MIDYukiko Kamiya0Yuuki Takeyama1Tomonari Mizuno2Fuminori Satoh3Hiroyuki Asanuma4Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, JapanDepartment of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, JapanDepartment of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, JapanDepartment of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, JapanDepartment of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, JapanSmall interfering RNA (siRNA) has been recognized as a powerful gene-silencing tool. For therapeutic application, chemical modification is often required to improve the properties of siRNA, including its nuclease resistance, activity, off-target effects, and tissue distribution. Careful siRNA guide strand selection in the RNA-induced silencing complex (RISC) is important to increase the RNA interference (RNAi) activity as well as to reduce off-target effects. The passenger strand-mediated off-target activity was previously reduced and on-target activity was enhanced by substitution with acyclic artificial nucleic acid, namely serinol nucleic acid (SNA). In the present study, the reduction of off-target activity caused by the passenger strand was investigated by modifying siRNAs with SNA. The interactions of SNA-substituted mononucleotides, dinucleotides, and (2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO)-labeled double-stranded RNA (dsRNA) with the MID domain of the Argonaute 2 (AGO2) protein, which plays a pivotal role in strand selection by accommodation of the 5’-terminus of siRNA, were comprehensively analyzed. The obtained nuclear magnetic resonance (NMR) data revealed that AGO2-MID selectively bound to the guide strand of siRNA due to the inhibitory effect of the SNA backbone located at the 5’ end of the passenger strand.https://www.mdpi.com/1422-0067/21/15/5218siRNARNAiAGO2MIDserinol nucleic acidoff-target effect |
| spellingShingle | Yukiko Kamiya Yuuki Takeyama Tomonari Mizuno Fuminori Satoh Hiroyuki Asanuma Investigation of Strand-Selective Interaction of SNA-Modified siRNA with AGO2-MID International Journal of Molecular Sciences siRNA RNAi AGO2 MID serinol nucleic acid off-target effect |
| title | Investigation of Strand-Selective Interaction of SNA-Modified siRNA with AGO2-MID |
| title_full | Investigation of Strand-Selective Interaction of SNA-Modified siRNA with AGO2-MID |
| title_fullStr | Investigation of Strand-Selective Interaction of SNA-Modified siRNA with AGO2-MID |
| title_full_unstemmed | Investigation of Strand-Selective Interaction of SNA-Modified siRNA with AGO2-MID |
| title_short | Investigation of Strand-Selective Interaction of SNA-Modified siRNA with AGO2-MID |
| title_sort | investigation of strand selective interaction of sna modified sirna with ago2 mid |
| topic | siRNA RNAi AGO2 MID serinol nucleic acid off-target effect |
| url | https://www.mdpi.com/1422-0067/21/15/5218 |
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