Characterizing and controlling CRISPR repair outcomes in nondividing human cells

作者：Gokul N. Ramadoss, Samali J. Namaganda, Manasi M. Kumar, Jennifer Hamilton, Rohit Sharma, Karena G. Chow, Luke A. Workley, Bria L Macklin, Mengyuan Sun, Alvin Ha, Jiacheng Liu, Christof Fellmann, Hannah L. Watry, Philip H. Dierks, Rudra S. Bose, Julianne Jin, Barbara S. Perez, Cindy R. Sandoval Espinoza, Madeline Matia, Serena Huei-An Lu, Luke M. Judge, Brian R. Shy, André Nussenzweig, Britt Adamson, Niren Murthy, Jennifer A. Doudna, Martin Kampmann, Bruce R. Conklin · 发表于：Nature Communications · 年份：2025 · DOI：10.1038/s41467-025-66058-3 · 被引用次数：14 · 研究领域：CRISPR and Genetic Engineering、Genetic Neurodegenerative Diseases、Pluripotent Stem Cells Research

Genome editing is poised to revolutionize treatment of genetic diseases, but poor understanding and control of DNA repair outcomes hinders its therapeutic potential. DNA repair is especially understudied in nondividing cells like neurons, limiting the efficiency and precision of genome editing in many clinically relevant tissues. Here, we address this barrier by using induced pluripotent stem cells (iPSCs) and iPSC-derived neurons to examine how postmitotic human neurons repair Cas9-induced DNA damage. CRISPR editing outcomes differ dramatically in neurons compared to genetically identical dividing cells: neurons take longer to fully resolve this damage, and upregulate non-canonical DNA repair factors in the process. Manipulating this response with chemical or genetic perturbations allows us to direct DNA repair toward desired editing outcomes in nondividing human neurons, cardiomyocytes, and primary T cells. By studying DNA repair in clinically relevant cells, we reveal unforeseen challenges and opportunities for precise therapeutic editing.