Characterizing and controlling CRISPR repair outcomes in nondividing human cells
Abstract
Abstract 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.
Article Details
Authors (28)
Gokul N. Ramadoss
Samali J. Namaganda
Manasi M. Kumar
Jennifer R. Hamilton
Rohit Sharma
Karena G. Chow
Luke A. Workley
Bria L. Macklin
Mengyuan Sun
Alvin S. Ha
Jia-Cheng Liu
Christof Fellmann
Hannah L. Watry
Philip H. Dierks
Rudra S. Bose
Julianne Jin
Barbara S. Perez
Cindy R. Sandoval Espinoza
Madeline P. Matia
Serena H. Lu
Luke M. Judge
Brian R. Shy
André Nussenzweig
Britt Adamson
Niren Murthy
Jennifer A. Doudna
Martin Kampmann
Bruce R. Conklin