Analyzing Balanced Reduction/Oxidation Status in Liver Cells Under Extreme Stresses with Vector-Mediated Genome-editing

Julia Houston, Dr. Ed Schmidt, Microbiology and Immunology, Montana State University, Schmidt Laboratory, MSU, M&I Department, PO Box 173520, Bozeman MT 59717

This project aims to develop a vector-mediated genome-editing system for genetic analyses of how liver cells tolerate extreme stresses that can disrupt the normal processes cells use to maintain a balanced reduction/oxidation status (“redox homeostasis”). Whereas most organisms require the activities of either glutathione reductase, thioredoxin reductase, or both to sustain redox homeostasis, our laboratory showed that liver cells can use, alternatively, a third pathway fueled by catabolism of the essential amino acid methionine. To define how this pathway works, we are developing a system to genetically target candidate genes associated with this pathway in individual cells within the livers of live mice. My project generates control vectors for this system. Briefly, vectors encoding genome-editing cassettes for disrupting internal positive-, negative-, and conditional-control genes will be developed to lay the foundation for future pathway analyses. Positive controls will disrupt genes encoding activities essential in all cells, such as for transcription or translation. Delivery of one of these vectors to a subset of cells within a mouse liver should specifically cause those cells to die and disappear, along with the vector that targeted them, from livers of any genotype. Negative controls will target genes that are nonessential in all cells. In this case, we expect the vectors to persist in livers of all genotypes. Finally, as “conditional-controls,” we will target two genes, cystathionase and glutamate-cysteine ligase, that we know are essential for the methionine catabolism pathway, but nonessential in wild type (WT) cells. These vectors should persist in WT livers but be lost from the livers of mice dependent on methionine catabolism for redox homeostasis. This project will allow us to precisely define the strategies that will be used in future studies to define how methionine catabolism can sustain redox homeostasis during extreme stress conditions.

Additional Abstract Information

Presenter: Julia Houston

Institution: Montana State University Bozeman

Type: Poster

Subject: Biology

Status: Approved

Time and Location

Session: Poster 2
Date/Time: Mon 3:00pm-4:00pm
Session Number: 2604