Tolerance for mutations and chemical modifications in a siRNA
Open accessAbstract
Short interfering RNA (siRNA), the active agent of RNA interference, shows promise of becoming a valuable tool in both basic and clinical research. We explore the tolerance to mutations and chemical modifications in various parts of the two 21-nt strands of a siRNA targeting the blood clotting initiator Tissue Factor. The mutations were G/C transversions. The chemical modifications were 2'-O-methylation, 2'-O-allylation and phosphorothioates. We found that siRNA generally tolerated mutations in the 5' end, while the 3' end exhibited low tolerance. This observation may facilitate the design of siRNA for specific targeting of transcripts containing single nucleotide polymorphisms. We further demonstrate that in our system the single antisense strand of the wild-type siRNA is almost as effective as the siRNA duplex, while the corresponding methylated M2+4 version of the antisense had reduced activity. Most of the chemically modified versions tested had near-wild-type initial activity, while the long-term activity was increased for certain siRNA species. Our results may improve the design of siRNAs for in vivo experiments.
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Increasing the PS content of individual siRNA duplexes to 80% can limit RISC efficacy and increase toxicity.
“However, increasing the PS content of individual siRNA duplexes to 80% can limit RNA-induced silencing complex (RISC) efficacy 17, 18 and increase toxicity19, 20.”
From A divalent siRNA chemical scaffold for potent and sustained modulation of gene expression throughout the central nervous system (2019) · cites this paper as [19]
The Chemical modification section reports about 70% cell death with the most extensively phosphorothioated siRNA, supporting the toxicity part. The Discussion says shorter phosphorothioate stretches were well tolerated; our copy does not specify 80% PS content or show that PS modification limits RISC efficacy. (LLM, conf=0.86)
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- Authors
- Amarzguioui, M.
- Affiliations
- University of Oslo (NO)
- Corresponding author
- Amarzguioui, M.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2003-01-15
- Volume
- 31
- Issue
- 2
- Pages
- 589-595
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- PMC
- PMC140512
- OpenAlex
- W2159595055
- Cited by
- 534
- References
- 1
- Citations by year
- 2026: 6 · 2025: 5 · 2024: 9 · 2023: 12 · 2022: 10 · 2021: 11 · 2020: 11 · 2019: 15 · 2018: 10 · 2017: 10 · 2016: 10 · 2015: 19 · 2014: 20 · 2013: 18 · 2012: 32
- FWCI
- 17.96
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Nucleic Acids Research (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Kreftforeningen
- Norges Forskningsråd
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Keywords
- siRNA, RNA interference, tissue factor, point mutations, chemical modification, 2'-O-methylation, phosphorothioate, antisense strand, single nucleotide polymorphisms, siRNA design, siRNA duplexes, in vivo gene silencing
- MeSH terms
- Cell Line, Humans, Mutation, Oligoribonucleotides, RNA, Messenger, Thromboplastin, Transfection, RNA Interference, RNA, Small Interfering
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