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Selection and Characterization of DNA Aptamers Targeting All Four Serotypes of Dengue Viruses.

Chen HL, Hsiao WH, Lee HC, Wu SC, Cheng JW - PLoS ONE (2015)

Bottom Line: Dengue viruses (DENVs) are members of Flaviviridae family, which are associated with human disease.S15 was found to form a parallel quadruplex based on Quadfinder prediction, gel mobility assay and circular dichroism studies.Our result provides a new opportunity in the development of DNA aptamers against DENVs in the future.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biotechnology and Department of Life Science, National Tsing Hua University, Hsinchu 300, Taiwan.

ABSTRACT
Dengue viruses (DENVs) are members of Flaviviridae family, which are associated with human disease. The envelope (E) protein plays an important role in viral infection. However, there is no effective antibody for clinical treatment due to antibody dependent enhancement of infection. In this study, using Systematic Evolution of Ligands by Exponential Enrichment (SELEX), we demonstrated the first aptamer (S15) that can bind to DENV-2 envelop protein domain III (ED3) with a high binding affinity. S15 was found to form a parallel quadruplex based on Quadfinder prediction, gel mobility assay and circular dichroism studies. Both the quadruplex structure and the sequence on 5'-end were necessary for the binding activity of S15. NMR titration experiments indicated that S15 bound to a highly conserved loop between βA and βB strands of ED3. Moreover, S15 can neutralize the infections by all four serotypes of DENVs. Our result provides a new opportunity in the development of DNA aptamers against DENVs in the future.

No MeSH data available.


Related in: MedlinePlus

Neutralization activity of S15 against all four DENV.Following incubation with various concentrations of aptamer S15 (0–100 μM), DENV HAWAII, DENV-2 NGC, DENV-3 H-87, and DENV-4 814669 were further incubated with Vero E6 cells, respectively. The infection was determined by PRNT assay. Samples were measured in triplicate and means were shown with standard error.
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pone.0131240.g004: Neutralization activity of S15 against all four DENV.Following incubation with various concentrations of aptamer S15 (0–100 μM), DENV HAWAII, DENV-2 NGC, DENV-3 H-87, and DENV-4 814669 were further incubated with Vero E6 cells, respectively. The infection was determined by PRNT assay. Samples were measured in triplicate and means were shown with standard error.

Mentions: Aptamer S15 was able to recognize DENV-2 virion as well as ED3 (data not shown), although the epitope appeared to be poorly accessible in the native virus structure [37]. We performed a PRNT assay to determine the neutralization activity against DENVs. The results showed that the infection of DENV-2 NGC was completely neutralized by 100 μM S15 (Fig 4). The IC50 was 4.2 μM. Moreover, using structure-based sequence alignment we found that residues (Q316, H317 and G318) on the binding epitope for aptamer S15 are identical (Fig 5). We therefore tested the neutralization activity against DENV-1 HAWAII, DENV-3 H-87 and DENV-4 H241. The IC50 values of the other three dengue serotypes are in similar range with DENV-2 NGC (IC50-DENV-1 = 1.7 μM, IC50-DENV-3 = 5.2 μM, IC50-DENV-4 = 5.8 μM). However, only DENV-2 NGC can be completely neutralized by S15 at 100 μM.


Selection and Characterization of DNA Aptamers Targeting All Four Serotypes of Dengue Viruses.

Chen HL, Hsiao WH, Lee HC, Wu SC, Cheng JW - PLoS ONE (2015)

Neutralization activity of S15 against all four DENV.Following incubation with various concentrations of aptamer S15 (0–100 μM), DENV HAWAII, DENV-2 NGC, DENV-3 H-87, and DENV-4 814669 were further incubated with Vero E6 cells, respectively. The infection was determined by PRNT assay. Samples were measured in triplicate and means were shown with standard error.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4482433&req=5

pone.0131240.g004: Neutralization activity of S15 against all four DENV.Following incubation with various concentrations of aptamer S15 (0–100 μM), DENV HAWAII, DENV-2 NGC, DENV-3 H-87, and DENV-4 814669 were further incubated with Vero E6 cells, respectively. The infection was determined by PRNT assay. Samples were measured in triplicate and means were shown with standard error.
Mentions: Aptamer S15 was able to recognize DENV-2 virion as well as ED3 (data not shown), although the epitope appeared to be poorly accessible in the native virus structure [37]. We performed a PRNT assay to determine the neutralization activity against DENVs. The results showed that the infection of DENV-2 NGC was completely neutralized by 100 μM S15 (Fig 4). The IC50 was 4.2 μM. Moreover, using structure-based sequence alignment we found that residues (Q316, H317 and G318) on the binding epitope for aptamer S15 are identical (Fig 5). We therefore tested the neutralization activity against DENV-1 HAWAII, DENV-3 H-87 and DENV-4 H241. The IC50 values of the other three dengue serotypes are in similar range with DENV-2 NGC (IC50-DENV-1 = 1.7 μM, IC50-DENV-3 = 5.2 μM, IC50-DENV-4 = 5.8 μM). However, only DENV-2 NGC can be completely neutralized by S15 at 100 μM.

Bottom Line: Dengue viruses (DENVs) are members of Flaviviridae family, which are associated with human disease.S15 was found to form a parallel quadruplex based on Quadfinder prediction, gel mobility assay and circular dichroism studies.Our result provides a new opportunity in the development of DNA aptamers against DENVs in the future.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biotechnology and Department of Life Science, National Tsing Hua University, Hsinchu 300, Taiwan.

ABSTRACT
Dengue viruses (DENVs) are members of Flaviviridae family, which are associated with human disease. The envelope (E) protein plays an important role in viral infection. However, there is no effective antibody for clinical treatment due to antibody dependent enhancement of infection. In this study, using Systematic Evolution of Ligands by Exponential Enrichment (SELEX), we demonstrated the first aptamer (S15) that can bind to DENV-2 envelop protein domain III (ED3) with a high binding affinity. S15 was found to form a parallel quadruplex based on Quadfinder prediction, gel mobility assay and circular dichroism studies. Both the quadruplex structure and the sequence on 5'-end were necessary for the binding activity of S15. NMR titration experiments indicated that S15 bound to a highly conserved loop between βA and βB strands of ED3. Moreover, S15 can neutralize the infections by all four serotypes of DENVs. Our result provides a new opportunity in the development of DNA aptamers against DENVs in the future.

No MeSH data available.


Related in: MedlinePlus