Limits...
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

Aptamers exhibited no cytotoxic effects in vitro and in vivo.a DNA aptamers exhibited no cytotoxicity in cultured cells. The results for each group are presented as the mean ± SD of three independent experiments. b No pathological changes were found in liver and spleen tissues of aptamer-injected turtles, compared with normal control turtles. The scale bars indicate 20 μm
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig4: Aptamers exhibited no cytotoxic effects in vitro and in vivo.a DNA aptamers exhibited no cytotoxicity in cultured cells. The results for each group are presented as the mean ± SD of three independent experiments. b No pathological changes were found in liver and spleen tissues of aptamer-injected turtles, compared with normal control turtles. The scale bars indicate 20 μm

Mentions: We incubated cells with the selected aptamers and then evaluated cell viability by the MTT method. There were no significant differences between the control and experimental groups, even at aptamer concentrations up to 1000 nM, suggesting that the aptamers were not cytotoxic (Fig. 4a). Furthermore, no turtles in the aptamer-treated or control groups had died 10 days p.i.. There were no pathological changes were in the liver or spleen in aptamer-injected turtles compared with controls, indicating that the selected aptamers had no cytotoxic effects in vivo (Fig. 4b).Fig. 4


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)

Aptamers exhibited no cytotoxic effects in vitro and in vivo.a DNA aptamers exhibited no cytotoxicity in cultured cells. The results for each group are presented as the mean ± SD of three independent experiments. b No pathological changes were found in liver and spleen tissues of aptamer-injected turtles, compared with normal control turtles. The scale bars indicate 20 μm
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig4: Aptamers exhibited no cytotoxic effects in vitro and in vivo.a DNA aptamers exhibited no cytotoxicity in cultured cells. The results for each group are presented as the mean ± SD of three independent experiments. b No pathological changes were found in liver and spleen tissues of aptamer-injected turtles, compared with normal control turtles. The scale bars indicate 20 μm
Mentions: We incubated cells with the selected aptamers and then evaluated cell viability by the MTT method. There were no significant differences between the control and experimental groups, even at aptamer concentrations up to 1000 nM, suggesting that the aptamers were not cytotoxic (Fig. 4a). Furthermore, no turtles in the aptamer-treated or control groups had died 10 days p.i.. There were no pathological changes were in the liver or spleen in aptamer-injected turtles compared with controls, indicating that the selected aptamers had no cytotoxic effects in vivo (Fig. 4b).Fig. 4

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