bibtex.bib

@article{Cloarec-Ung_Beaulieu_Suthananthan_Lehnertz_Sauvageau_Sheppard_Knapp_2024,
	title        = {
		Near-perfect precise on-target editing of human hematopoietic stem and
		progenitor cells
	},
	author       = {
		Cloarec-Ung, Fanny-Mei and Beaulieu, Jamie and Suthananthan, Arunan and
		Lehnertz, Bernhard and Sauvageau, Guy and Sheppard, Hilary M. and Knapp,
		David J. H. F.
	},
	year         = 2024,
	month        = jun,
	journal      = {eLife},
	volume       = 12,
	pages        = {RP91288},
	doi          = {10.7554/eLife.91288},
	issn         = {2050-084X},
	abstractnote = {
		Precision gene editing in primary hematopoietic stem and progenitor cells
		(HSPCs) would facilitate both curative treatments for monogenic disorders as
		well as disease modelling. Precise efficiencies even with the CRISPR/Cas
		system, however, remain limited. Through an optimization of guide RNA
		delivery, donor design, and additives, we have now obtained mean precise
		editing efficiencies >90% on primary cord blood HSCPs with minimal toxicity
		and without observed off-target editing. The main protocol modifications
		needed to achieve such high efficiencies were the addition of the DNA-PK
		inhibitor AZD7648, and the inclusion of spacer-breaking silent mutations in
		the donor in addition to mutations disrupting the PAM sequence. Critically,
		editing was even across the progenitor hierarchy, did not substantially
		distort the hierarchy or affect lineage outputs in colony-forming cell assays
		or the frequency of high self-renewal potential long-term culture initiating
		cells. As modelling of many diseases requires heterozygosity, we also
		demonstrated that the overall editing and zygosity can be tuned by adding in
		defined mixtures of mutant and wild-type donors. With these optimizations,
		editing at near-perfect efficiency can now be accomplished directly in human
		HSPCs. This will open new avenues in both therapeutic strategies and disease
		modelling.
	},
	language     = {eng}
}