美国麻省理工学院Francisco J. Sánchez Rivera研究组利用先导编辑传感器文库高通量评估遗传变异。该项研究成果于2024年3月12日在线发表在《自然—生物技术》杂志上。

据悉，肿瘤基因组通常包含复杂的单核苷酸改变和染色体重排，这些改变和重排会扰乱蛋白质的功能。先导编辑已被用于安装和评估遗传变异，但以前的方法一直受限于先导编辑向导RNA的不同效率。

附：英文原文

Title: High-throughput evaluation of genetic variants with prime editing sensor libraries

Author: Gould, Samuel I., Wuest, Alexandra N., Dong, Kexin, Johnson, Grace A., Hsu, Alvin, Narendra, Varun K., Atwa, Ondine, Levine, Stuart S., Liu, David R., Snchez Rivera, Francisco J.

Issue&Volume: 2024-03-12

Abstract: Tumor genomes often harbor a complex spectrum of single nucleotide alterations and chromosomal rearrangements that can perturb protein function. Prime editing has been applied to install and evaluate genetic variants, but previous approaches have been limited by the variable efficiency of prime editing guide RNAs. Here we present a high-throughput prime editing sensor strategy that couples prime editing guide RNAs with synthetic versions of their cognate target sites to quantitatively assess the functional impact of endogenous genetic variants. We screen over 1,000 endogenous cancer-associated variants of TP53—the most frequently mutated gene in cancer—to identify alleles that impact p53 function in mechanistically diverse ways. We find that certain endogenous TP53 variants, particularly those in the p53 oligomerization domain, display opposite phenotypes in exogenous overexpression systems. Our results emphasize the physiological importance of gene dosage in shaping native protein stoichiometry and protein–protein interactions, and establish a framework for studying genetic variants in their endogenous sequence context at scale.

DOI: 10.1038/s41587-024-02172-9

Source: https://www.nature.com/articles/s41587-024-02172-9