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ARS Home » Southeast Area » Stoneville, Mississippi » Genomics and Bioinformatics Research » Research » Research Project #443104

Research Project: The Development of Novel RNA Dependent RNA Polymerases, Nucleases, and Transposases by Metagenomics and Protein Engineering

Location: Genomics and Bioinformatics Research

Project Number: 6066-21310-006-023-S
Project Type: Non-Assistance Cooperative Agreement

Start Date: Sep 1, 2022
End Date: Aug 30, 2024

1. We will use metagenomic approaches to discover novel enzymes that amplify RNA or modify DNA. These enzymes will be synthesized and characterized to evaluate their potential use in biotechnology and diagnostics. Initially, we will identify RNA-dependent RNA polymerases (RdRPs) with high fidelity. Current siagnostic tests for RNA detection use and DNA intermediate using enzymes like reverse transcriptase that slows the testing. The discovery of high-fidelity RdRPs could enable the direct detection of RNA and open up a new class of molecular diagnostics. 2. Expand these metagenomic approaches to other enzymes related to CRISPR detection like transposes and Cas enzymes.

Viruses use RNA-dependent RNA polymerases (RdRPs) to copy their RNA genomes. Most RNA viruses lack error correction capabilities and this limits the length of their genomes to about 10,000 bases. We will search metagenomics data for long contigs containing RdRPs then synthesize the RdRPs and measure their error rate and reaction kinetics.