Neuronal DNA repair reveals strategies to influence CRISPR editing outcomes.

Autor:	Ramadoss GN; Gladstone Institutes, San Francisco, CA, 94158, USA.; Institute for Neurodegenerative Diseases, University of California, San Francisco, CA, 94158, USA., Namaganda SJ; Gladstone Institutes, San Francisco, CA, 94158, USA., Hamilton JR; Innovative Genomics Institute, University of California, Berkeley, CA, 94720, USA.; Department of Molecular & Cell Biology, University of California, Berkeley, CA, 94720, USA., Sharma R; Innovative Genomics Institute, University of California, Berkeley, CA, 94720, USA.; Department of Bioengineering, University of California, Berkeley, CA, 94720, USA., Chow KG; Gladstone Institutes, San Francisco, CA, 94158, USA., Macklin BL; Gladstone Institutes, San Francisco, CA, 94158, USA., Sun M; Gladstone Institutes, San Francisco, CA, 94158, USA., Liu JC; Laboratory of Genome Integrity, National Cancer Institute, NIH, Bethesda, MD, 20892, USA., Fellmann C; Gladstone Institutes, San Francisco, CA, 94158, USA.; Department of Cellular & Molecular Pharmacology, University of California, San Francisco, CA, 94158, USA., Watry HL; Gladstone Institutes, San Francisco, CA, 94158, USA., Jin J; Institute for Neurodegenerative Diseases, University of California, San Francisco, CA, 94158, USA., Perez BS; Innovative Genomics Institute, University of California, Berkeley, CA, 94720, USA.; Department of Molecular & Cell Biology, University of California, Berkeley, CA, 94720, USA., Sandoval Espinoza CR; Innovative Genomics Institute, University of California, Berkeley, CA, 94720, USA.; Department of Molecular & Cell Biology, University of California, Berkeley, CA, 94720, USA., Matia MP; Gladstone Institutes, San Francisco, CA, 94158, USA., Lu SH; Gladstone Institutes, San Francisco, CA, 94158, USA., Judge LM; Gladstone Institutes, San Francisco, CA, 94158, USA.; Department of Pediatrics, University of California, San Francisco, CA, 94158, USA., Nussenzweig A; Laboratory of Genome Integrity, National Cancer Institute, NIH, Bethesda, MD, 20892, USA., Adamson B; Department of Molecular Biology, Princeton University, Princeton, NJ, 08544, USA.; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, 08544, USA., Murthy N; Innovative Genomics Institute, University of California, Berkeley, CA, 94720, USA.; Department of Bioengineering, University of California, Berkeley, CA, 94720, USA., Doudna JA; Gladstone Institutes, San Francisco, CA, 94158, USA.; Innovative Genomics Institute, University of California, Berkeley, CA, 94720, USA.; Department of Molecular & Cell Biology, University of California, Berkeley, CA, 94720, USA.; California Institute for Quantitative Biosciences, University of California, Berkeley, CA, 94720, USA.; Howard Hughes Medical Institute, University of California, Berkeley, CA, 94720, USA.; Department of Chemistry, University of California, Berkeley, CA, 94720, USA.; MBIB Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA., Kampmann M; Institute for Neurodegenerative Diseases, University of California, San Francisco, CA, 94158, USA.; Department of Biochemistry & Biophysics, University of California, San Francisco, CA, 94158, USA., Conklin BR; Gladstone Institutes, San Francisco, CA, 94158, USA.; Innovative Genomics Institute, University of California, Berkeley, CA, 94720, USA.; Department of Cellular & Molecular Pharmacology, University of California, San Francisco, CA, 94158, USA.; Department of Medicine, University of California, San Francisco, CA, 94158, USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Jun 26. Date of Electronic Publication: 2024 Jun 26.
DOI:	10.1101/2024.06.25.600517
Abstrakt:	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, which must withstand decades of DNA damage without replicating. This lack of knowledge limits the efficiency and precision of genome editing in clinically relevant cells. To address this, we used induced pluripotent stem cells (iPSCs) and iPSC-derived neurons to examine how postmitotic human neurons repair Cas9-induced DNA damage. We discovered that neurons can take weeks to fully resolve this damage, compared to just days in isogenic iPSCs. Furthermore, Cas9-treated neurons upregulated unexpected DNA repair genes, including factors canonically associated with replication. Manipulating this response with chemical or genetic perturbations allowed us to direct neuronal repair toward desired editing outcomes. By studying DNA repair in postmitotic human cells, we uncovered unforeseen challenges and opportunities for precise therapeutic editing. Competing Interests: DECLARATION OF INTERESTS MK is a co-scientific founder of Montara Therapeutics and serves on the Scientific Advisory Boards of Engine Biosciences, Casma Therapeutics, Cajal Neuroscience, Alector, and Montara Therapeutics, and is an advisor to Modulo Bio and Recursion Therapeutics. MK is an inventor on US Patent 11,254,933 related to CRISPRi and CRISPRa screening, and on a US Patent application on in vivo screening methods. JRH is a co-founder of Azalea Therapeutics. CF is a co-founder of Mirimus, Inc. B.A. is an advisory board member with options for Arbor Biotechnologies and Tessera Therapeutics. BA holds equity in Celsius Therapeutics. The Regents of the University of California have patents issued and pending for CRISPR technologies (on which JAD is an inventor) and delivery technologies (on which JAD and JRH are co-inventors). JAD is a cofounder of Azalea Therapeutics, Caribou Biosciences, Editas Medicine, Evercrisp, Scribe Therapeutics, Intellia Therapeutics, and Mammoth Biosciences. JAD is a scientific advisory board member at Evercrisp, Caribou Biosciences, Intellia Therapeutics, Scribe Therapeutics, Mammoth Biosciences, The Column Group and Inari. JAD is Chief Science Advisor to Sixth Street, a Director at Johnson & Johnson, Altos and Tempus, and has a research project sponsored by Apple Tree Partners.
Databáze:	MEDLINE
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