Genome Engineering of Hematopoietic Stem Cells Using CRISPR/Cas9 System.
Title | Genome Engineering of Hematopoietic Stem Cells Using CRISPR/Cas9 System. |
Publication Type | Journal Article |
Year of Publication | 2022 |
Authors | Devaraju N, Rajendiran V, Ravi NSam, Mohankumar KM |
Journal | Methods Mol Biol |
Volume | 2429 |
Pagination | 307-331 |
Date Published | 2022 |
ISSN | 1940-6029 |
Keywords | Animals, CRISPR-Cas Systems, Gene Editing, Hematopoietic Stem Cell Transplantation, Hematopoietic Stem Cells, Mice, Transplantation, Autologous |
Abstract | 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. |
DOI | 10.1007/978-1-0716-1979-7_20 |
Alternate Journal | Methods Mol Biol |
PubMed ID | 35507170 |