Effect of 2'-5'/3'-5' phosphodiester linkage heterogeneity on RNA interference.
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
21 05 2020
21 05 2020
Historique:
accepted:
25
03
2020
revised:
19
03
2020
received:
20
02
2020
pubmed:
14
4
2020
medline:
9
9
2020
entrez:
14
4
2020
Statut:
ppublish
Résumé
We report on the synthesis of siRNAs containing both 2'-5'- and 3'-5'-internucleotide linkages and their effects on siRNA structure, function, and interaction with RNAi proteins. Screening of these siRNAs against their corresponding mRNA targets showed that 2'-5' linkages were well tolerated in the sense strand, but only at a few positions in the antisense strand. Extensive modification of the antisense strand minimally affected 5'-phosphorylation of the siRNA by kinases, however, it negatively affected siRNA loading into human AGO2. Modelling and molecular dynamics simulations were fully consistent with these findings. Furthermore, our studies indicated that the presence of a single 5'p-rN1-(2'-5')-N2 unit in the antisense strand does not alter the 'clover leaf' bend and sugar puckers that are critical for anchoring the 5'-phosphate to Ago 2 MID domain. Importantly, 2'-5'-linkages had the added benefit of abrogating immune-stimulatory activity of siRNAs. Together, these results demonstrate that 2'-5'/3'-5'-modified siRNAs, when properly designed, can offer an efficient new class of siRNAs with diminished immune-stimulatory responses.
Identifiants
pubmed: 32282904
pii: 5819601
doi: 10.1093/nar/gkaa222
pmc: PMC7229817
doi:
Substances chimiques
AGO2 protein, human
0
Argonaute Proteins
0
RNA, Small Interfering
0
Tumor Suppressor Protein p53
0
Luciferases, Firefly
EC 1.13.12.7
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
4643-4657Subventions
Organisme : NIAID NIH HHS
ID : R01 AI042552
Pays : United States
Organisme : CIHR
ID : FDN-148366
Pays : Canada
Informations de copyright
© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.
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