@article {2545,
	title = {Genome Engineering of Hematopoietic Stem Cells Using CRISPR/Cas9 System.},
	journal = {Methods Mol Biol},
	volume = {2429},
	year = {2022},
	month = {2022},
	pages = {307-331},
	abstract = {<p>Ex vivo genetic manipulation of autologous hematopoietic stem and progenitor cells (HSPCs) is a viable strategy for the treatment of hematologic and primary immune disorders. Targeted genome editing of HSPCs using the CRISPR-Cas9 system provides an effective platform to edit the desired genomic locus for therapeutic purposes with minimal off-target effects. In this chapter, we describe the detailed methodology for the CRISPR-Cas9 mediated gene knockout, deletion, addition, and correction in human HSPCs by viral and nonviral approaches. We also present a comprehensive protocol for the analysis of genome modified HSPCs toward the erythroid and megakaryocyte lineage in vitro and the long-term multilineage reconstitution capacity in the recently developed NBSGW mouse model that supports human erythropoiesis.</p>
},
	keywords = {Animals, CRISPR-Cas Systems, Gene Editing, Hematopoietic Stem Cell Transplantation, Hematopoietic Stem Cells, Mice, Transplantation, Autologous},
	issn = {1940-6029},
	doi = {10.1007/978-1-0716-1979-7_20},
	author = {Devaraju, Nivedhitha and Rajendiran, Vignesh and Ravi, Nithin Sam and Mohankumar, Kumarasamypet M}
}
