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 |
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. See: https://www.partybuslimosoc.com/corporate
CRISPR/Cas9 is a powerful gene cutting and editing tool that allows researchers to interfere with genes by cutting, modifying, or replacing specific sections of genes. https://thepasswordgame.io
Revolutionizing Therapeutic Potential. Explore how CRISPR/Cas9 technology enables precise modifications to hematopoietic stem cells, promising breakthroughs in treating genetic blood disorders and advancing personalized medicine."
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Genome engineering of hematopoietic stem cells (HSCs) using CRISPR/Cas9 offers potential for treating genetic blood disorders. The CRISPR system, comprising Cas9 enzyme and guide RNA, enables precise DNA modification. HSCs are isolated, cultured, and CRISPR components are delivered via viral vectors or electroporation. Look it up here https://savvytipsguru.com/tips-on-how-to-work-two-part-time-jobs/.
Guide RNAs are designed to target specific DNA sequences, inducing breaks. Natural repair mechanisms either create mutations (NHEJ) or incorporate desired changes (HDR). Edited HSCs are screened and transplanted back into the patient, potentially correcting genetic anomalies. Challenges include off-target effects, efficient delivery, and safety. Ongoing research aims to refine this method for effective and secure genome editing in HSCs. https://answersville.com/.
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