Development of RNA aptamer that inhibits methyltransferase activity of dengue virus
To develop an RNA aptamer specific for the methyltransferase (MTase) of dengue virus (DENV) which is essential for viral genome replication and translation acting directly on N-7 and 2′-O-methylation of the type-I cap structure of the viral RNA.
We identified 2′-fluoro-modified RNA aptamers that can specifically bind DENV serotype 2 (DENV2) MTase using systematic evolution of ligands by exponential enrichment technology. We truncated the chosen aptamer into a 45-mer RNA sequence that can bind DENV2 MTase with K d ~ 28 nM and inhibit N-7 methylation activity of the protein. Moreover, the 45-mer truncated aptamer could not only bind with an K d ~ 15.6 nM but also inhibit methylation activity of DENV serotype 3 (DENV3) MTase. The 45-mer aptamer competitively impeded binding of both DENV2 and DENV3 genomic RNA to MTase of each serotype.
The selected 45-mer truncated RNA aptamer specifically and avidly bound DENV MTase and competitively inhibited its methylation activity, and thus could be useful for the development of anti-DENV agents.
KeywordsAntiviral agent Dengue virus Methyltransferase RNA aptamer
The present research was conducted by the research fund of Dankook University in Republic of Korea in 2015.
Supplementary Table 1—Sequence of primers.
Supplementary Fig. 1—Enrichment of selected RNA aptamer pool.
Supplementary Fig. 2—Aptamer truncation experiment.
Supplementary Fig. 3—Comparison of amino acid sequence of MTase protein of DENV 1-4 serotypes.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
- Kim SH, Yang IY, Jang SH, Kim J, Truong TT, Van Pham T, Truong NU, Lee KY, Jang YS (2013) C5a receptor-targeting ligand-mediated delivery of dengue virus antigen to M cells evokes antigen-specific systemic and mucosal immune responses in oral immunization. Microbes Infect 15:895–902CrossRefPubMedGoogle Scholar