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Evaluation of multiple-turnover capability of locked nucleic acid antisense oligonucleotides in cell-free RNase H-mediated antisense reaction and in mice.
- Source :
-
Nucleic acid therapeutics [Nucleic Acid Ther] 2014 Aug; Vol. 24 (4), pp. 283-90. Date of Electronic Publication: 2014 Apr 23. - Publication Year :
- 2014
-
Abstract
- The multiple-turnover ability of a series of locked nucleic acid (LNA)-based antisense oligonucleotides (AONs) in the RNase H-mediated scission reaction was estimated using a newly developed cell-free reaction system. We determined the initial reaction rates of AONs under multiple-turnover conditions and found that among 24 AONs tested, AONs with melting temperatures (Tm) of 40°C-60°C efficiently elicit multiple rounds of RNA scission. On the other hand, by measuring Tm with two 10-mer RNAs partially complementary to AONs as models of cleaved 5' and 3' fragments of mRNA, we found that AONs require adequate binding affinity for efficient turnover activities. We further demonstrated that the efficacy of a set of 13-mer AONs in mice correlated with their turnover efficiency, indicating that the intracellular situation where AONs function is similar to multiple-turnover conditions. Our methodology and findings may provide an opportunity to shed light on a previously unknown antisense mechanism, leading to further improvement of the activity and safety profiles of AONs.
- Subjects :
- Animals
Apolipoprotein B-100 antagonists & inhibitors
Apolipoprotein B-100 metabolism
Cell-Free System
Gene Expression
Half-Life
Injections, Subcutaneous
Liver chemistry
Liver metabolism
Male
Mice
Mice, Inbred C57BL
Oligonucleotides metabolism
Oligonucleotides pharmacokinetics
Oligonucleotides, Antisense metabolism
Oligonucleotides, Antisense pharmacokinetics
RNA Stability
RNA, Messenger antagonists & inhibitors
RNA, Messenger metabolism
Ribonuclease H metabolism
Transition Temperature
Apolipoprotein B-100 genetics
Oligonucleotides chemistry
Oligonucleotides, Antisense chemistry
RNA, Messenger genetics
Ribonuclease H chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 2159-3345
- Volume :
- 24
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Nucleic acid therapeutics
- Publication Type :
- Academic Journal
- Accession number :
- 24758560
- Full Text :
- https://doi.org/10.1089/nat.2013.0470