CRISPRtOE gives scientists a leg up on bacteria

Tool for bacterial gene overexpression could enable breakthroughs in medicine, bioproducts

A team of scientists from UW-Madison, including Jason Peters, has developed a tool that researchers can use to better understand bacteria and their genes in the pursuit of better antibiotic drugs and microbes used in bioproduct manufacturing.

A man in a white lab coat and blue latex gloves pipettes liquid under a ventilation hood. — WEI affiliate Jason Peters, an associate professor of medical genetics at UW–Madison, develops toolkits that help researchers identify key genes in bacteria, which can lead to breakthroughs in antibiotic treatments and bioproduct manufacturing. Image by: Sally Griffith-Oh/UW–Madison

CRISPRtOE is the latest CRISPR-based tool developed by the Peters Lab. They tested the tool, called CRISPR Transposition and OverExpression (CRISPRtOE), in Escherichia coli K-12, a non-pathogenic model organism used to study fundamental biological processes, and Zymomonas mobilis, a microbe that’s good at making biofuels but less well understood. CRISPRtOE allows scientists studying bacteria to insert promoter genes that boost the activity of nearby genes, which can help pinpoint the genes that help the organism resist toxins encountered while manufacturing bioproducts or delivered in the form of antibiotic drug treatments.

A man smiles while operating a touchscreen gel electrophoresis instrument in a lab, with shelves of lab equipment and a whiteboard visible in the background — Ryan Ward, a graduate student in the Peters Lab and contributor to the CRISPRtOE project, zooms in on a gel electrophoresis reading used for DNA analysis. Image by: Chelsea Mamott/Wisconsin Energy Institute

Peter anticipates CRISPRtOE can be readily applied to other microbes and screening conditions. The scalable approaches of CRISPRtOE can be adapted to diverse bacteria, a vital step in bridging the gap between genome sequencing and gene function assessment.

“That CRISPRtOE can target all genes in a bacterium means we can understand the cellular roles of ‘quiet’ genes that are normally turned off,” Peters said. “The ever-expanding world of bacterial genetics will always keep us on our toes!”

Read the full article at: https://energy.wisc.edu/news/crisprtoe-gives-scientists-leg-bacteria?fbclid=IwY2xjawU8sSlleHRuA2FlbQIxMABwZG9mBWJyaWQRMWQzUVo1SGo0ZFdOTmNScmtzcnRjBmFwcF9pZBAyMjIwMzkxNzg4MjAwODkyAAEe0sO_6D5P6glu0GZi_Yygi54vjpxJalu9lbKptgBfszhZa6y1RZoVCNFt03g_aem_7umrwGtIOC6Awp_iGSAwRw