Journal article
Transient inhibition of 53BP1 increases the frequency of targeted integration in human hematopoietic stem and progenitor cells
Nature communications, Vol.15(1), pp.111-14
01/02/2024
DOI: 10.1038/s41467-023-43413-w
PMCID: PMC10762240
PMID: 38169468
Abstract
Genome editing by homology directed repair (HDR) is leveraged to precisely modify the genome of therapeutically relevant hematopoietic stem and progenitor cells (HSPCs). Here, we present a new approach to increasing the frequency of HDR in human HSPCs by the delivery of an inhibitor of 53BP1 (named “i53”) as a recombinant peptide. We show that the use of i53 peptide effectively increases the frequency of HDR-mediated genome editing at a variety of therapeutically relevant loci in HSPCs as well as other primary human cell types. We show that incorporating the use of i53 recombinant protein allows high frequencies of HDR while lowering the amounts of AAV6 needed by 8-fold. HDR edited HSPCs were capable of long-term and bi-lineage hematopoietic reconstitution in NSG mice, suggesting that i53 recombinant protein might be safely integrated into the standard CRISPR/AAV6-mediated genome editing protocol to gain greater numbers of edited cells for transplantation of clinically meaningful cell populations.
Here the authors demonstrate that the frequency of HDR in human hematopoietic stem and progenitor cells is increased by the delivery of an inhibitor of 53BP1 as a recombinant peptide. This approach is applicable for a variety of therapeutically relevant loci in HSPCs as well in other primary human cell types.
Details
- Title: Subtitle
- Transient inhibition of 53BP1 increases the frequency of targeted integration in human hematopoietic stem and progenitor cells
- Creators
- Ron Baik - Stanford University School of MedicineM. Kyle Cromer - Stanford University School of MedicineSteve E. Glenn - Integrated DNA Technologies (United States)Christopher A. Vakulskas - Integrated DNA Technologies (United States)Kay O. Chmielewski - University of FreiburgAmanda M. Dudek - Stanford University School of MedicineWilliam N. Feist - Stanford University School of MedicineJulia Klermund - University of FreiburgSuzette Shipp - Stanford University School of MedicineToni Cathomen - University of FreiburgDaniel P. Dever - Stanford University School of MedicineMatthew H. Porteus - Stanford University School of Medicine
- Resource Type
- Journal article
- Publication Details
- Nature communications, Vol.15(1), pp.111-14
- DOI
- 10.1038/s41467-023-43413-w
- PMID
- 38169468
- PMCID
- PMC10762240
- NLM abbreviation
- Nat Commun
- ISSN
- 2041-1723
- eISSN
- 2041-1723
- Publisher
- Nature Publishing Group UK
- Language
- English
- Date published
- 01/02/2024
- Academic Unit
- Microbiology and Immunology
- Record Identifier
- 9984822991902771
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