Using CRISPRi to Repress Cell’s Ability to Metabolize Glycerol and to Express LacZ

Abstract

CRISPR interference (CRISPRi) method is an effective way in disturbing a specific gene in the DNA by blocking transcription and therefore silencing a gene from expressing.  When the catalytically dead Cas9 is coexpressed with a guide RNA, it generates a DNA recognition complex that can target a specific gene. The project focuses on manipulating the genes of DNA to repress certain gene expressions.  The first assay involves repressing the genes that allow cells to grow in glycerol, and the second assay targets the cell's ability to break down lacZ.