Deep CRISPR mutagenesis characterizes the functional diversity of TP53 mutations
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Open Access
- 7 January 2025
- journal article
- research article
- Published by Springer Nature in Nature Genetics
- Vol. 57 (1) , 140-153
Abstract
The mutational landscape of TP53 , a tumor suppressor mutated in about half of all cancers, includes over 2,000 known missense mutations. To fully leverage TP53 mutation status for personalized medicine, a thorough understanding of the functional diversity of these mutations is essential. We conducted a deep mutational scan using saturation genome editing with CRISPR-mediated homology-directed repair to engineer 9,225 TP53 variants in cancer cells. This high-resolution approach, covering 94.5% of all cancer-associated TP53 missense mutations, precisely mapped the impact of individual mutations on tumor cell fitness, surpassing previous deep mutational scan studies in distinguishing benign from pathogenic variants. Our results revealed even subtle loss-of-function phenotypes and identified promising mutants for pharmacological reactivation. Moreover, we uncovered the roles of splicing alterations and nonsense-mediated messenger RNA decay in mutation-driven TP53 dysfunction. These findings underscore the power of saturation genome editing in advancing clinical TP53 variant interpretation for genetic counseling and personalized cancer therapy.Keywords
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Funding Information
- Deutsche Forschungsgemeinschaft (STI 182/15-1)
- Deutsche Forschungsgemeinschaft (STI 182/13-1)
- Deutsche Forschungsgemeinschaft (GRK2573)
- Deutsche Forschungsgemeinschaft (109546710 Project A10)
- Deutsche Forschungsgemeinschaft (GRK2573)
- Deutsche Forschungsgemeinschaft (SA 1923/7-1)
- Deutsche Forschungsgemeinschaft (CRC1213 A10N)
- Deutsche Forschungsgemeinschaft (JO 1473/1-3)
- Bundesministerium für Forschung, Technologie und Raumfahrt (031L0063)
- Bundesministerium für Forschung, Technologie und Raumfahrt (DZL)
- Hessisches Ministerium für Wissenschaft und Kunst (LOEWE iCANx)
- Von-Behring-Röntgen-Stiftung (65-0004)
- Von-Behring-Röntgen-Stiftung (66-LV06)
- Von-Behring-Röntgen-Stiftung (71-0012)
- José Carreras Leukämie-Stiftung (09 R/2018)
- Structural Genomics Consortium (SGC): https://www.thesgc.org
- Institute of Lung Health
- Deutsche Krebshilfe (TACTIC)
This publication has 97 references indexed in Scilit:
- Gain of Function of a p53 Hot Spot Mutation in a Mouse Model of Li-Fraumeni SyndromeCell, 2004
- Chromosome stability, in the absence of apoptosis, is critical for suppression of tumorigenesis in Trp53 mutant miceNature Genetics, 2003
- Understanding the function–structure and function–mutation relationships of p53 tumor suppressor protein by high-resolution missense mutation analysisProceedings of the National Academy of Sciences, 2003
- Restoration of the tumor suppressor function to mutant p53 by a low-molecular-weight compoundNature Medicine, 2002
- Rescuing the function of mutant p53Nature Reviews Cancer, 2001
- PUMA, a Novel Proapoptotic Gene, Is Induced by p53Molecular Cell, 2001
- Requirement for p53 and p21 to Sustain G 2 Arrest After DNA DamageScience, 1998
- p53 levels, functional domains, and DNA damage determine the extent of the apoptotic response of tumor cells.Genes & Development, 1996
- Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumoursNature, 1992
- Germ Line p53 Mutations in a Familial Syndrome of Breast Cancer, Sarcomas, and Other NeoplasmsScience, 1990