Functional screening in human HSPCs identifies optimized protein-based enhancers of Homology Directed Repair

Homology Directed Repair (HDR) enables precise genome editing, but the implementation of HDR-based therapies is hindered by limited efficiency in comparison to methods that exploit alternative DNA repair routes, such as Non-Homologous End Joining (NHEJ). In this study, we develop a functional, poole...

Full description

Saved in:

Bibliographic Details
Published in:	Nature communications Vol. 15; no. 1; p. 2625
Main Authors:	Perez-Bermejo, Juan A., Efagene, Oghene, Matern, William M., Holden, Jeffrey K., Kabir, Shaheen, Chew, Glen M., Andreoletti, Gaia, Catton, Eniola, Ennis, Craig L., Garcia, Angelica, Gerstenberg, Trevor L., Hill, Kaisle A., Jain, Aayami, Krassovsky, Kristina, Lalisan, Cassandra D., Lord, Daniel, Quejarro, B. Joy, Sales-Lee, Jade, Shah, Meet, Silva, Brian J., Skowronski, Jason, Strukov, Yuri G., Thomas, Joshua, Veraz, Michael, Vijay, Twaritha, Wallace, Kirby A., Yuan, Yue, Grogan, Jane L., Wienert, Beeke, Lahiri, Premanjali, Treusch, Sebastian, Dever, Daniel P., Soros, Vanessa B., Partridge, James R., Seim, Kristen L.
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 23-03-2024 Nature Publishing Group Nature Portfolio
Subjects:	13/31 45/100 45/23 45/77 631/1647/2163 631/337/1427/2190 631/61/201/2110 82/47 82/75 Additives DNA repair Editing Enhancers Gene therapy Genetic modification Genome editing Genomes Hematopoietic stem cells Hemopoiesis Homologous recombination Homology Humanities and Social Sciences Inhibitors multidisciplinary Non-homologous end joining Progenitor cells Proteins Reagents Science Science (multidisciplinary) Screening Stem cells Synergism
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	Homology Directed Repair (HDR) enables precise genome editing, but the implementation of HDR-based therapies is hindered by limited efficiency in comparison to methods that exploit alternative DNA repair routes, such as Non-Homologous End Joining (NHEJ). In this study, we develop a functional, pooled screening platform to identify protein-based reagents that improve HDR in human hematopoietic stem and progenitor cells (HSPCs). We leverage this screening platform to explore sequence diversity at the binding interface of the NHEJ inhibitor i53 and its target, 53BP1, identifying optimized variants that enable new intermolecular bonds and robustly increase HDR. We show that these variants specifically reduce insertion-deletion outcomes without increasing off-target editing, synergize with a DNAPK inhibitor molecule, and can be applied at manufacturing scale to increase the fraction of cells bearing repaired alleles. This screening platform can enable the discovery of future gene editing reagents that improve HDR outcomes. Here the authors describe a functional screening platform in human stem cells to identify and optimize protein-based gene editing additives that increase homologous directed recombination and have potential to improve gene therapy workflows.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	2041-1723 2041-1723
DOI:	10.1038/s41467-024-46816-5