麻省理工学院和哈佛大学的布罗德研究所John G. Doench小组近日取得一项新成果。经过不懈努力，他们开发出用于增强碱基编辑器突变扫描的基于活动的选择。这一研究成果于2025年10月14日发表在国际顶尖学术期刊《自然—遗传学》上。

在这里，研究小组开发了一种共选择方法，丰富了具有高碱基编辑活性的细胞，大大提高了目标位点的编辑效率。该课题组对这种基于活性的选择方法进行了评估，对比传统的筛选方法，通过在TP53上平纹引导RNAs，证明了其精确定位特定突变和功能重要的蛋白质区域的能力增强。该课题组研究人员预计这种模块化选择方法将提高许多应用程序中碱基编辑屏幕的分辨率。

据了解，碱基编辑是一种基于CRISPR的技术，可以对基因组进行高通量、核苷酸水平的功能查询，这对于理解人类疾病的遗传基础和为治疗开发提供信息至关重要。碱基编辑屏幕已经成为一种强大的实验方法，然而，编辑效率在细胞间的显著差异引入了噪声，可能会模糊有意义的结果。

附：英文原文

Title: Activity-based selection for enhanced base editor mutational scanning

Author: Kaplan, Eleanor G., Steger, Ryan J., Shah, Spencer T., Drepanos, Laura M., Griffith, Audrey L., Reint, Ganna, Doench, John G.

Issue&Volume: 2025-10-14

Abstract: Base editing is a CRISPR-based technology that enables high-throughput, nucleotide-level functional interrogation of the genome that is essential for understanding the genetic basis of human disease and informing therapeutic development. Base editing screens have emerged as a powerful experimental approach, yet significant cell-to-cell variability in editing efficiency introduces noise that may obscure meaningful results. Here we develop a co-selection method that enriches for cells with high base editing activity, substantially increasing editing efficiency at a target locus. We evaluate this activity-based selection method against a traditional screening approach by tiling guide RNAs across TP53, demonstrating its enhanced capacity to pinpoint specific mutations and protein regions of functional importance. We anticipate that this modular selection method will enhance the resolution of base editing screens across many applications.

DOI: 10.1038/s41588-025-02366-0

Source: https://www.nature.com/articles/s41588-025-02366-0