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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

Sequence and frequency of selected aptamers.Selected aptamers were sequenced and were classified into 7 groups.
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pone.0131240.g001: Sequence and frequency of selected aptamers.Selected aptamers were sequenced and were classified into 7 groups.

Mentions: A single-strained DNA library containing approximately 6×1014 molecules was used to select DNA aptamers against DENV-2 ED3. After 15 rounds of selection, we obtained twelve different sequences from fifty clones. According to their sequences, these aptamers were classified into groups A-G (Fig 1). Group A (44%) and group B (28%) were dominant. In contrast, sequences in groups C-G showed much lower frequency. Two dominant aptamer, S22 (frequency = 19/50) and S15 (frequency = 12/50), were selected for the following studies.


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)

Sequence and frequency of selected aptamers.Selected aptamers were sequenced and were classified into 7 groups.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0131240.g001: Sequence and frequency of selected aptamers.Selected aptamers were sequenced and were classified into 7 groups.
Mentions: A single-strained DNA library containing approximately 6×1014 molecules was used to select DNA aptamers against DENV-2 ED3. After 15 rounds of selection, we obtained twelve different sequences from fifty clones. According to their sequences, these aptamers were classified into groups A-G (Fig 1). Group A (44%) and group B (28%) were dominant. In contrast, sequences in groups C-G showed much lower frequency. Two dominant aptamer, S22 (frequency = 19/50) and S15 (frequency = 12/50), were selected for the following studies.

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