瑞士苏黎世联邦理工学院Randall J. Platt研究小组，使用碱基编辑和先导编辑实现基因变异的多模态扫描。这一研究成果于2024年11月12日在线发表在国际学术期刊《自然—生物技术》上。

研究人员结合了细胞嘧啶和腺嘌呤碱基编辑器以及先导编辑器，评估了表皮生长因子受体基因（EGFR）中广泛变异的致病性。

通过使用合成的碱基编辑和原位编辑导向RNA库，研究人员在多个细胞系中安装了数万个涵盖EGFR全编码序列的变异，并评估这些变异在肿瘤发生和耐酪氨酸激酶抑制剂中的作用。

EGFR变异扫描识别出重要的结果，验证了该方法的鲁棒性，并揭示了EGFR激活和药物反应中未被充分认识的路径。

研究人员预期，多模态精确突变扫描可以广泛应用于任何感兴趣的基因元素的遗传变异表征，并在高分辨率和单核苷酸级别上进行分析。

据介绍，突变扫描将基因变异与表型联系起来，使得能够探讨蛋白质功能、相互作用和变异致病性。然而，当前的方法在高通量地工程化定制化的、多样化的基因变异集合，尤其是在内源性基因座和不同细胞背景下，效率较低。

附：英文原文

Title: Multimodal scanning of genetic variants with base and prime editing

Author: Belli, Olivier, Karava, Kyriaki, Farouni, Rick, Platt, Randall J.

Issue&Volume: 2024-11-12

Abstract: Mutational scanning connects genetic variants to phenotype, enabling the interrogation of protein functions, interactions and variant pathogenicity. However, current methodologies cannot efficiently engineer customizable sets of diverse genetic variants in endogenous loci across cellular contexts in high throughput. Here, we combine cytosine and adenine base editors and a prime editor to assess the pathogenicity of a broad spectrum of variants in the epithelial growth factor receptor gene (EGFR). Using pooled base editing and prime editing guide RNA libraries, we install tens of thousands of variants spanning the full coding sequence of EGFR in multiple cell lines and assess the role of these variants in tumorigenesis and resistance to tyrosine kinase inhibitors. Our EGFR variant scan identifies important hits, supporting the robustness of the approach and revealing underappreciated routes to EGFR activation and drug response. We anticipate that multimodal precision mutational scanning can be applied broadly to characterize genetic variation in any genetic element of interest at high and single-nucleotide resolution.

DOI: 10.1038/s41587-024-02439-1

Source: https://www.nature.com/articles/s41587-024-02439-1