德国马尔堡大学Thorsten Stiewe等研究人员合作发现，深度CRISPR诱变技术表征TP53突变的功能多样性。相关论文于2025年1月7日在线发表在《自然—遗传学》杂志上。

研究人员通过使用CRISPR介导的同源定向修复进行饱和基因组编辑，开展了深度诱变扫描，成功地在癌细胞中构建了9225种TP53变异。这种高分辨率的方法覆盖了94.5%的所有与癌症相关的TP53错义突变，精确地描绘了个别突变对肿瘤细胞适应性的影响，超越了以往的深度诱变扫描研究，在区分良性突变和致病性突变方面取得了显著进展。

结果揭示了即使是细微的功能丧失表型，并确定了有前景的突变体用于药理学重激活。此外，研究人员还揭示了剪接改变和无义介导的信使RNA降解在突变驱动的TP53功能失常中的作用。这些发现强调了饱和基因组编辑在推动临床TP53变异解释方面的强大潜力，并为遗传咨询和个性化癌症治疗提供了重要的见解。

据悉，TP53的突变景观是一个在约一半癌症中出现的肿瘤抑制基因，其中包括2000多个已知的错义突变。为了充分利用TP53突变状态进行个性化医学治疗，深入了解这些突变的功能多样性至关重要。

附：英文原文

Title: Deep CRISPR mutagenesis characterizes the functional diversity of TP53 mutations

Author: Funk, Julianne S., Klimovich, Maria, Drangenstein, Daniel, Pielhoop, Ole, Hunold, Pascal, Borowek, Anna, Noeparast, Maxim, Pavlakis, Evangelos, Neumann, Michelle, Balourdas, Dimitrios-Ilias, Kochhan, Katharina, Merle, Nastasja, Bullwinkel, Imke, Wanzel, Michael, Elmshuser, Sabrina, Teply-Szymanski, Julia, Nist, Andrea, Procida, Tara, Bartkuhn, Marek, Humpert, Katharina, Mernberger, Marco, Savai, Rajkumar, Soussi, Thierry, Joerger, Andreas C., Stiewe, Thorsten

Issue&Volume: 2025-01-07

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.

DOI: 10.1038/s41588-024-02039-4

Source: https://www.nature.com/articles/s41588-024-02039-4