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.


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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.

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Tsz-Leung To, Ph.D.

Dr. To is currently a Senior Research Scientist and Group Leader in the Mootha laboratory and Metabolism Program at Broad Institute. He received his PhD in chemical engineering at Massachusetts Institute of Technology, where he focused on systems and quantitative biology. After spending a short stint in a private biotech company working on synthetic biology and metabolic engineering, he subsequently pursued his postdoctoral fellowship at UCSF where he focused on assay development and protein design. Dr. To has deep expertise in mitochondrial bioenergetics and specifically in massively parallel and pooled genetic screening, and he has led studies on systematic dissection of human mitochondria using genome-wide CRISPR.

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