Prime editing enables high-throughput identification and characterization of selective MutSβ inhibitors
Year: 2026
Type: preprint
Abstract: ABSTRACT DNA mismatch repair (MMR) maintains genome stability but paradoxically drives expansion of pathogenic repeats responsible for trinucleotide repeat disorders. Attenuating repeat instability through targeted inhibition of MMR is therefore a therapeutic strategy of interest; however, progress is constrained by a lack of robust methods for defining and isolating MMR subpathways. We exploited features of prime editing to profile the effects of core MMR genes on thousands of programmed DNA mispairs in cells and resolved distinct MMR processes, including a noncanonical “loop-directed” mechanism. Guided by these results, we built a set of scalable reporters that detect loss of separable MMR processes and screened ∼72,000 small molecules for activity against MutSβ-directed MMR. This effort identified a novel series of MutSβ inhibitors that reduced (CAG) n expansion in a cell model. Altogether, this study establishes several methods for systematically interrogating MMR, with demonstrated capacity to identify inhibitors of therapeutic value.
Authors: Yangwode Jing, Jun Yan, Denny Yang, Yanjin Chen, Issam Senoussi, James Woodward, Katya Williams, Hahn Kim, Kangning He, Herwin Daub, Vinay Dahiya, Maren Thomsen, Jia Ling, Purnima Ravisankar, Yuri Pritykin, Gabriel Balmus, Petr Cejka, Jeffrey A. Hussmann, Longbin Liu, Thomas F. Vogt, Michael Finley, Britt Adamson
Institutions: Princeton University, Gladstone Institutes, University of California, San Francisco, University of Cambridge, UK Dementia Research Institute, ETH Zurich, Institute for Research in Biomedicine, Institute of Biochemistry, Università della Svizzera italiana, Cornell University, Ludwig Cancer Research, Weill Cornell Medicine, Transylvanian Institute of Neuroscience, CHDI Foundation
Language: English
Topic: DNA Repair Mechanisms
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Funders: CHDI Foundation, Princeton University, Rutgers Cancer Institute of New Jersey, Ludwig Institute for Cancer Research, China Scholarship Council, UK Dementia Research Institute, National Institutes of Health, Medical Research Council, National Human Genome Research Institute, National Cancer Institute, National Institute of General Medical Sciences
Awards: G2104897622, G2131311388, G3664767099, G3670273158, G6309704273, Mapping the DNA damage response in human cells with high-resolution functional genomics, G9000033417, Cancer Center Support Grant, Center for Genomic Editing and Recording: Development and Application of Next-Generation Genome and Epigenome Editing Methods to Advance the Study and Treatment of Human Disease, Flow Cytometry and Cell Sorting
OpenAlex ID: W7220697173