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Characterization of DNA aptamers generated against the soft-shelled turtle iridovirus with antiviral effects.

Li P, Zhou L, Yu Y, Yang M, Ni S, Wei S, Qin Q - BMC Vet. Res. (2015)

Bottom Line: Soft-shelled turtle iridovirus (STIV) causes severe systemic disease in farmed soft-shelled turtles (Trionyx sinensis).Electrophoretic mobility shift assays and fluorescent localization showed that the selected aptamers had high binding affinity for STIV.Aptamer QA-36 had the highest calculated binding affinity (K d ) of 53.8 nM.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou, 510301, China.

ABSTRACT

Background: Soft-shelled turtle iridovirus (STIV) causes severe systemic disease in farmed soft-shelled turtles (Trionyx sinensis). More efficient methods of controlling and detecting STIV infections are urgently needed. 

Methods: In this study, we generated eight single-stranded DNA (ssDNA) aptamers against STIV using systematic evolution of ligands by exponential enrichment (SELEX).

Results: The aptamers formed representative stem-loop secondary structures. Electrophoretic mobility shift assays and fluorescent localization showed that the selected aptamers had high binding affinity for STIV. Aptamer QA-36 had the highest calculated binding affinity (K d ) of 53.8 nM. Flow cytometry and fluorescence microscopy of cell-aptamer interactions demonstrated that QA-12 was able to recognize both STIV-infected cells and tissues with a high level of specificity. Moreover, the selected aptamers inhibited STIV infection in vitro and in vivo, with aptamer QA-36 demonstrating the greatest protective effect against STIV and inhibiting STIV infection in a dose-dependent manner.

Discussion: We generated DNA aptamers that bound STIV with a high level of specificity, providing an alternative means for investigating STIV pathogenesis, drug development, and medical therapies for STIV infection.

Conclusions: These DNA aptamers may thus be suitable antiviral candidates for the control of STIV infections.

No MeSH data available.


Related in: MedlinePlus

Selected aptamers inhibited STIV infection in cultured cells. a Morphology of FHM cells following treatment with the SELEX library or selected aptamers. b Cell morphology and CPE after treatment with STIV previously incubated with or without selected aptamers. c Aptamers reduced virus titers in cultured cells. d QA-36 aptamer inhibited STIV infection in a dose-dependent manner. The results for each group are presented as the mean ± SD of three independent experiments. (**P < 0.01; *P < 0.05; bars = 50 μm)
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Fig5: Selected aptamers inhibited STIV infection in cultured cells. a Morphology of FHM cells following treatment with the SELEX library or selected aptamers. b Cell morphology and CPE after treatment with STIV previously incubated with or without selected aptamers. c Aptamers reduced virus titers in cultured cells. d QA-36 aptamer inhibited STIV infection in a dose-dependent manner. The results for each group are presented as the mean ± SD of three independent experiments. (**P < 0.01; *P < 0.05; bars = 50 μm)

Mentions: Cells without any treatment were served as mock group and grew normally. Cells incubated with SELEX library or aptamers also maintained normal growth, it indicated that selected aptamers exhibited no cytotoxic effects in cell cultures, which was consistent with the results of Fig. 4. (Fig. 5a). The inhibitory effects of aptamers was shown in Fig. 5b. There were significant CPEs in control cells incubated with STIV alone, when our aptamers were added, although there were some CPEs, the CPEs were much less than the control groups, which means STIV infection could be partially inhibited by the selected aptamers (Fig. 5b). Virus titers were significantly reduced by all the aptamers at 48 h post-infection, with QA-36 displaying the greatest inhibitory effect on STIV among the tested aptamers (Fig. 5c).Fig. 5


Characterization of DNA aptamers generated against the soft-shelled turtle iridovirus with antiviral effects.

Li P, Zhou L, Yu Y, Yang M, Ni S, Wei S, Qin Q - BMC Vet. Res. (2015)

Selected aptamers inhibited STIV infection in cultured cells. a Morphology of FHM cells following treatment with the SELEX library or selected aptamers. b Cell morphology and CPE after treatment with STIV previously incubated with or without selected aptamers. c Aptamers reduced virus titers in cultured cells. d QA-36 aptamer inhibited STIV infection in a dose-dependent manner. The results for each group are presented as the mean ± SD of three independent experiments. (**P < 0.01; *P < 0.05; bars = 50 μm)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4588899&req=5

Fig5: Selected aptamers inhibited STIV infection in cultured cells. a Morphology of FHM cells following treatment with the SELEX library or selected aptamers. b Cell morphology and CPE after treatment with STIV previously incubated with or without selected aptamers. c Aptamers reduced virus titers in cultured cells. d QA-36 aptamer inhibited STIV infection in a dose-dependent manner. The results for each group are presented as the mean ± SD of three independent experiments. (**P < 0.01; *P < 0.05; bars = 50 μm)
Mentions: Cells without any treatment were served as mock group and grew normally. Cells incubated with SELEX library or aptamers also maintained normal growth, it indicated that selected aptamers exhibited no cytotoxic effects in cell cultures, which was consistent with the results of Fig. 4. (Fig. 5a). The inhibitory effects of aptamers was shown in Fig. 5b. There were significant CPEs in control cells incubated with STIV alone, when our aptamers were added, although there were some CPEs, the CPEs were much less than the control groups, which means STIV infection could be partially inhibited by the selected aptamers (Fig. 5b). Virus titers were significantly reduced by all the aptamers at 48 h post-infection, with QA-36 displaying the greatest inhibitory effect on STIV among the tested aptamers (Fig. 5c).Fig. 5

Bottom Line: Soft-shelled turtle iridovirus (STIV) causes severe systemic disease in farmed soft-shelled turtles (Trionyx sinensis).Electrophoretic mobility shift assays and fluorescent localization showed that the selected aptamers had high binding affinity for STIV.Aptamer QA-36 had the highest calculated binding affinity (K d ) of 53.8 nM.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou, 510301, China.

ABSTRACT

Background: Soft-shelled turtle iridovirus (STIV) causes severe systemic disease in farmed soft-shelled turtles (Trionyx sinensis). More efficient methods of controlling and detecting STIV infections are urgently needed. 

Methods: In this study, we generated eight single-stranded DNA (ssDNA) aptamers against STIV using systematic evolution of ligands by exponential enrichment (SELEX).

Results: The aptamers formed representative stem-loop secondary structures. Electrophoretic mobility shift assays and fluorescent localization showed that the selected aptamers had high binding affinity for STIV. Aptamer QA-36 had the highest calculated binding affinity (K d ) of 53.8 nM. Flow cytometry and fluorescence microscopy of cell-aptamer interactions demonstrated that QA-12 was able to recognize both STIV-infected cells and tissues with a high level of specificity. Moreover, the selected aptamers inhibited STIV infection in vitro and in vivo, with aptamer QA-36 demonstrating the greatest protective effect against STIV and inhibiting STIV infection in a dose-dependent manner.

Discussion: We generated DNA aptamers that bound STIV with a high level of specificity, providing an alternative means for investigating STIV pathogenesis, drug development, and medical therapies for STIV infection.

Conclusions: These DNA aptamers may thus be suitable antiviral candidates for the control of STIV infections.

No MeSH data available.


Related in: MedlinePlus