A variant-to-function map of POLG via deep mutational scanning
Lay Abstract
POLG deficiency is associated with tremendous allelic heterogeneity and pleiotropy, hence making it very challenging to know which observed variants are benign versus pathogenic. Our goal is to create a “look up table” for POLG that connects all possible coding variants to function. We will leverage next generation DNA synthesis to create a saturation mutagenesis library for POLG and then perform pooled functional genetic screens that quantitatively score POLG variants based on their impact on mitochondrial oxidative phosphorylation. The proposed project takes advantage of our lab’s longstanding commitment to mitochondrial disease, expertise in pooled genetic screening for mitochondrial function, as well as the Broad Institute’s proven expertise in mammalian genetic screening. Our goal is to create a durable resource that will be useful to the research and clinical communities.
Institution
Broad Institute of MIT and Harvard
415 Main Street
Cambridge, MA 0214
Principle Investigators
Vamsi K. Mootha, M.D.
Dr. Vamsi Mootha is currently an Investigator of the Howard Hughes Medical Institute, Professor of Systems Biology and Medicine at Harvard Medical School, Investigator in the Department of Molecular Biology at Massachusetts General Hospital and founding Co-Director of the Broad Institute’s Metabolism Program. Dr. Mootha is an internationally recognized physician-scientist and leader in mitochondrial physiology and disease for which he received a MacArthur Prize, the King Faisal Prize in Science, a Padma Shri from the Government of India, and election to both the National Academy of Sciences and National Academy of Medicine. His team characterized the mitochondrial proteome, discovered the calcium uniporter, identified twenty Mendelian disease genes, and discovered that in animal models low oxygen can alleviate mitochondrial disease. His laboratory has deep expertise in the application of modern genomics and genetics methods to investigate mitochondrial disease biology.
See Full Bio Here >
Anna Kotrys, Ph.D.
Anna received her B.S in biotechnology and M.S in molecular biology from the University of Warsaw. During her undergraduate studies she received the Rector’s Scholarship for outstanding undergraduate work. She then completed her PhD at the Institute of Biochemistry and Biophysics in Warsaw under mentorship of Dr. Roman Szczesny focusing on the mitochondrial nucleic acids biology. Her work was awarded with the Prime Minister of Poland award and recognition from Merck. In 2020, Anna started her postdoctoral work in the laboratory of Prof. Vamsi Mootha. She was a part of the research team to report the first successful, precise editing of mtDNA. She also developed a novel targeted, modular and scalable single-cell sequencing technology. Between 2021-2024 Anna was supported by HHMI Jane Coffin Childs Fellowship.