Precise measurement of CRISPR genome editing outcomes through single-cell DNA sequencing

作者：Nechama Kalter, Saurabh Gulati, Michael A. Rosenberg, Qawer Ayaz, Joanne Nguyen, Shu Wang, Benjamin Schroeder, Chieh-Yuan Li, Ayal Hendel · 发表于：Molecular Therapy — Methods & Clinical Development · 年份：2025 · DOI：10.1016/j.omtm.2025.101449 · 被引用次数：8 · 研究领域：CRISPR and Genetic Engineering、Innovation and Socioeconomic Development、Mosquito-borne diseases and control

Gene therapy for clinical applications necessitates a comprehensive, accurate, and precise measurement of gene-edited drug products. State-of-the-art pipelines for evaluating editing outcomes rely primarily on bulk sequencing approaches, which are limited to population-level assessment. Here, we leveraged Tapestri, a single-cell sequencing technology for an in-depth analysis of editing outcomes. Using this platform, we characterized the genotype of triple-edited cells simultaneously at more than 100 loci, including editing zygosity, structural variations, and cell clonality. Our findings revealed a unique editing pattern in nearly every edited cell, highlighting the importance of single-cell resolution measurement to ensure the highest safety standards.