SEARCH FOR A PSYCHROTOLERANT RNA POLYMERASE PROMOTER-SEQUENCE FOR ITS USE IN RIBOSWITCH BIOSENSORS
Abstract
Biosensor systems are gaining relevance due to their versatility in a broad spectrum of applications. They have revolutionized the field of medical diagnostics by providing fast and accurate sensors for different biological and chemical markers. Rapid Diagnostics Tests (RDTs), optimized for hostile environments, are as well an evident need. Accurate biosensors for the identification of pathogens may allow timely treatment and fast disease control, minimizing negative effects on general population. Most assay components, including enzymes, used by RDTs are designed for state-of-the-art facilities with proper storing and handling conditions. Therefore, for RDT’s that can be used in remote or complex areas with limited resources, require robust assay components and minimal or no refrigeration as key enabling elements. A biosensor is a self-contained, integrated receptor–transducer device capable of providing quantitative or semi-quantitative analytical information. Riboswitches or RNA-based intracellular gene expression switches, offer a way to synthetically develop biosensors to any analyte of interest. The design of these cell-free systems based on extremophilic enzymes has been explored to enable field applications, as they can retain their function and perform reactions in extreme environments. We have been able to detect and partially purified (80% purity) a novel RNA polymerase from an Antarctic virus which is highly active, being an extremely interesting protein for riboswitch implementation. Nevertheless, the RNA polymerase found during our previous AFOSR project grant and its promoter sequence, remains unknown. To know the exact sequence of the promoter that is recognized for the RNA polymerase is crucial for a biosensor application, as the promoter is the specific nucleotide sequence where the RNA polymerase interacts and binds to initiate the transcription of a gene, synthetizing a new RNA molecule from the DNA template.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Aug 12, 2021
- Source ID
- FA95502010390
Entities
People
- Jenny M Blamey
Organizations
- Air Force Office of Scientific Research
- United States Air Force