Dnazyme-based antiviral technology for agricultural crop protection

Overview

This solution leverages the 10-23 DNAzyme, a catalytic single-stranded DNA sequence, to combat agriculturally relevant viral diseases. DNAzymes can be designed to cleave virtually any RNA sequence between a purine and pyrimidine nucleotide pair by flanking a catalytic core with two substrate-specific binding arms. The approach aims to provide a curative treatment for viral infections in economically important crops, starting with maize dwarf mosaic virus (MDMV).

The technology builds on validated expertise in designing DNAzymes against the human prion protein mRNA, using bioinformatic methods that account for target site accessibility and product dissociation. This same design framework will be applied to identify optimal cleavage sites within plant viral RNA genomes, enabling precise and effective antiviral intervention.

Technical specifications

Core technology:

• 10-23 DNAzyme catalytic core flanked by two customizable binding arms
• Bioinformatic-driven design pipeline that evaluates cleavage site accessibility within full-length viral RNA
• Prediction of DNAzyme:RNA complex specificity, stability, and cleavage product dissociation
• Validated using electrophoretic mobility shift assays and Förster resonance energy transfer-based kinetic measurements

Design criteria for agricultural targets:

• Identification of suitable RY cleavage sites in viral RNA (e.g., MDMV)
• Assessment of secondary structure to ensure target accessibility
• Evaluation of binding arm specificity and complex stability
• Testing against short synthetic RNA sequences and full-length viral RNA transcripts

Key advantages:

• Programmable design applicable to diverse viral RNA targets
• Catalytic mechanism enabling multiple cleavage cycles per DNAzyme molecule
• Sequence-specific cleavage reducing off-target effects

Technology readiness level

The DNAzyme platform has been validated for cleaving synthetic and in vitro-transcribed RNA targets, with demonstrated efficiency in kinetic and binding assays. The design methodology, including bioinformatic site selection and activity assessment, is established and transferable to new viral targets. Current development focuses on extending the validated approach to agriculturally relevant viruses, with initial work targeting maize dwarf mosaic virus. The next validation phase includes bioinformatic analysis of MDMV RNA, commercial synthesis of candidate DNAzymes, and activity testing against short and full-length viral RNA sequences.