Aptamer-targeting of Aleutian mink disease virus (AMDV) can be an effective strategy to inhibit virus replication

Hui Zhang,Shuguang Wu,Hongyan Chen,Taofeng Lu,Lili Zhao,Qin Ma,Jie Zhou,Wenzhuo Yan

doi:10.1038/s41598-021-84223-8

Abstract

Aleutian mink disease (AMD), which is caused by Aleutian mink disease virus (AMDV), is an important contagious disease for which no effective vaccine is yet available. AMD causes major economic losses for mink farmers globally and threatens some carnivores such as skunks, genets, foxes and raccoons. Aptamers have exciting potential for the diagnosis and/or treatment of infectious viral diseases, including AMD. Using a magnetic beads-based systemic evolution of ligands by exponential enrichment (SELEX) approach, we have developed aptamers with activity against AMDV after 10 rounds of selection. After incubation with the ADVa012 aptamer (4 μM) for 48 h, the concentration of AMDV in the supernatant of infected cells was 47% lower than in the supernatant of untreated cells, whereas a random library of aptamers has no effect. The half-life of ADVa012 was ~ 32 h, which is significantly longer than that of other aptamers. Sequences and three dimensions structural modeling of selected aptamers indicated that they fold into similar stem-loop structures, which may be a preferred structure for binding to the target protein. The ADVa012 aptamer was shown to have an effective and long-lasting inhibitory effect on viral production in vitro.

Highlights

Aleutian mink disease (AMD), which is caused by Aleutian mink disease virus (AMDV), is an important contagious disease for which no effective vaccine is yet available
The magnetic beads coupled to recombinant AMDV VP2 protein can be used for target selection
If the plateau phase of the quantitative real-time PCR (qPCR) curves from last few rounds tends to be stable, we can assume that the diversity of oligonucleotides was exponentially reducing during the systemic evolution of ligands by exponential enrichment (SELEX) process and that the screening process was successful

Summary

Introduction

Aleutian mink disease (AMD), which is caused by Aleutian mink disease virus (AMDV), is an important contagious disease for which no effective vaccine is yet available. Aptamers have exciting potential for the diagnosis and/or treatment of infectious viral diseases, including AMD. Aleutian mink disease (AMD), which is caused by Aleutian mink disease virus (AMDV), is an important contagious disease that causes major economic losses for mink farmers g lobally[1]. Aptamers for various targets, including proteins, peptides16, bacteria17, viruses18,19, cells[20] and small molecules[21], have been selected in vitro using the systematic evolution of ligands by exponential enrichment strategy (SELEX). We obtained aptamers for AMDV VP2 protein using in vitro magnetic beads-based SELEX, and evaluated the antiviral activity of the selected aptamers in an AMDV infection assay.

Methods

Results

Conclusion