Ali Haidar
Pronouns: He/Him
Research Mentor(s): Shelby Hemker
Research Mentor School/College/Department: Human Genetics / Medicine
Program: UROPF
Session: Session 3 (11:00am – 11:50am)
Authors: Shelby Hemker, Ali Haidar
Abstract
Loss of functional alleles of the DNA repair gene MUTYHare linked to hereditary colorectal cancer risk syndromes. MUTYHis responsible for repairing 8oxo-guanine (8oxoG) to adenine mispairs, thus an accumulation of this DNA damage in the genome increases risk for cancer. However in the absence of MUTYH, 8oxoG:A mispairs can be repaired. In yeast, the mismatch repair (MMR) complex MutS-alpha is used to initiate this repair. The aim of this study is to determine if MMR can repair 8oxoG:A mispairs in humans. CRISPR/Cas9 genome editing was used to create frameshift mutations in the two MMR genes that make up MutS-alpha: MSH2 and MSH6. This was done in both wildtype HEK293 cells and in cells with deleterious mutations in MUTYHand OGG1, which together restore 8oxoG:A mispairs to G:C base pairs. The ability of these cell lines to repair 8oxoG:A mispairs will be tested with a novel GFP-off reporter assay. In this assay, GFP expression is turned on if 8xoxG:A mispairs are repaired. Thus, if the MUTYH, OGG1, and MMR triple knockout cell lines repair less GFP-off than the MUTYH, OGG1 double knockout cell lines, then MMR is capable of repairing 8oxoG:A mispairs.
