An Application of Mycobacteriophage Genome Engineering using Bacteriophage Recombineering with Electroporated DNA (BRED) and CRISPR Cas-9 Systems
Subject Area
Chemistry
Description
Ethan Dionne ’22
Majors: Biochemistry and Biology
Faculty Mentor: Dr. Kathleen Cornely, Chemistry and Biochemistry
The increasing occurrence of antibiotic-resistant pathogens remains a global threat to public health, spurring research to develop alternative treatments for infectious bacteria. Phage therapy, the targeted use of bacteria-specific viral particles called bacteriophages, has emerged as a promising antibiotic alternative. For the use of some phage in therapeutic applications, gene deletions are often necessary to ensure bacterial host lethality. Here we present an application of the CRISPY-BRED technique for the deletion of the immunity repressor gene in the mycobacteriophage known as Mufasa. This process involves a recombination event coupled with a CRISPR Cas-9 gene knockdown system for selection against the parent phage. With host cell transformation of this CRISPR Cas9 plasmid and inhibition of overall parent phage gene expression, straightforward mutant recovery should be achievable.
Publisher
Providence College
Date
4-27-2022
Type
Poster
Language
English
