Nonviral Ex Vivo Genome Editing in Mouse Bona Fide Hematopoietic Stem Cells with CRISPR/Cas9

Hiromasa Hara; Natsagdorj Munkh-Erdene; Suvd Byambaa; Yutaka Hanazono

doi:10.1007/978-1-0716-3016-7_16

Nonviral Ex Vivo Genome Editing in Mouse Bona Fide Hematopoietic Stem Cells with CRISPR/Cas9

Methods Mol Biol. 2023:2637:213-221. doi: 10.1007/978-1-0716-3016-7_16.

Authors

Hiromasa Hara^{1

2}, Natsagdorj Munkh-Erdene¹, Suvd Byambaa¹, Yutaka Hanazono^{3

4}

Affiliations

¹ Division of Regenerative Medicine, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan.
² Division of Development of Animal Resource, Center for Development of Advanced Medical Technology, Jichi Medical University, Shimotsuke, Tochigi, Japan.
³ Division of Regenerative Medicine, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan. hanazono@jichi.ac.jp.
⁴ Division of Development of Animal Resource, Center for Development of Advanced Medical Technology, Jichi Medical University, Shimotsuke, Tochigi, Japan. hanazono@jichi.ac.jp.

PMID: 36773149
DOI: 10.1007/978-1-0716-3016-7_16

Abstract

Knock-in therapy, in which an insertion site can be controlled, would be more suitable for the treatment of genetic blood disorders as compared to conventional gene therapy with lentivirus vectors that introduce genes into the genome randomly. Recent advancements in genome editing technology have substantially improved the knock-in efficiency, making it a reality. We present the details of a virus-free CRISPR/Cas9-based genome editing method for bona fide mouse hematopoietic stem cells.

Keywords: CRISPR/Cas9; Genome editing; Knock-in; Mouse hematopoietic stem cells; Plasmid.

MeSH terms

Animals
CRISPR-Cas Systems*
Gene Editing* / methods
Genetic Therapy / methods
Lentivirus / genetics
Mice