Overcoming GNA/RNA base-pairing limitations using isonucleotides improves the pharmacodynamic activity of ESC+ GalNAc-siRNAs.
Acetylgalactosamine
Adenosine
/ chemistry
Alcohol Oxidoreductases
/ antagonists & inhibitors
Animals
Base Pairing
COS Cells
Chlorocebus aethiops
Cytidine
/ chemistry
Dimethylformamide
/ analogs & derivatives
Ethylamines
/ chemistry
Female
Glycols
/ chemistry
Guanosine
/ chemistry
Hepatocytes
/ cytology
Hydrogen Bonding
Mice
Mice, Inbred C57BL
Oligoribonucleotides
/ chemistry
Organophosphorus Compounds
/ chemistry
Prealbumin
/ antagonists & inhibitors
Primary Cell Culture
RNA Stability
RNA, Double-Stranded
/ chemistry
RNA, Small Interfering
/ chemistry
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
08 11 2021
08 11 2021
Historique:
accepted:
27
09
2021
revised:
19
09
2021
received:
30
03
2021
pubmed:
15
10
2021
medline:
24
12
2021
entrez:
14
10
2021
Statut:
ppublish
Résumé
We recently reported that RNAi-mediated off-target effects are important drivers of the hepatotoxicity observed for a subset of GalNAc-siRNA conjugates in rodents, and that these findings could be mitigated by seed-pairing destabilization using a single GNA nucleotide placed within the seed region of the guide strand. Here, we report further investigation of the unique and poorly understood GNA/RNA cross-pairing behavior to better inform GNA-containing siRNA design. A reexamination of published GNA homoduplex crystal structures, along with a novel structure containing a single (S)-GNA-A residue in duplex RNA, indicated that GNA nucleotides universally adopt a rotated nucleobase orientation within all duplex contexts. Such an orientation strongly affects GNA-C and GNA-G but not GNA-A or GNA-T pairing in GNA/RNA heteroduplexes. Transposition of the hydrogen-bond donor/acceptor pairs using the novel (S)-GNA-isocytidine and -isoguanosine nucleotides could rescue productive base-pairing with the complementary G or C ribonucleotides, respectively. GalNAc-siRNAs containing these GNA isonucleotides showed an improved in vitro activity, a similar improvement in off-target profile, and maintained in vivo activity and guide strand liver levels more consistent with the parent siRNAs than those modified with isomeric GNA-C or -G, thereby expanding our toolbox for the design of siRNAs with minimized off-target activity.
Identifiants
pubmed: 34648028
pii: 6396890
doi: 10.1093/nar/gkab916
pmc: PMC8565336
doi:
Substances chimiques
Ethylamines
0
Glycols
0
Oligoribonucleotides
0
Organophosphorus Compounds
0
Prealbumin
0
RNA, Double-Stranded
0
RNA, Small Interfering
0
Ttr protein, mouse
0
phosphoramidite
0
Guanosine
12133JR80S
isoguanosine
1818-71-9
dimethylformamide-dimethylacetal
4637-24-5
isocytidine
489-59-8
Cytidine
5CSZ8459RP
N,N-diisopropylethylamine
5SIQ15721L
Dimethylformamide
8696NH0Y2X
Alcohol Oxidoreductases
EC 1.1.-
glycollate oxidase
EC 1.1.3.15
Adenosine
K72T3FS567
Acetylgalactosamine
KM15WK8O5T
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
10851-10867Informations de copyright
© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.
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