CRISPR/Cas9 Gene Targeting in Primary Mouse Bone Marrow-Derived Macrophages

Will Bailis

doi:10.1007/978-1-0716-0203-4_14

CRISPR/Cas9 Gene Targeting in Primary Mouse Bone Marrow-Derived Macrophages

Methods Mol Biol. 2020:2097:223-230. doi: 10.1007/978-1-0716-0203-4_14.

Author

Will Bailis¹

Affiliation

¹ Division for Protective Immunity, Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia, Philadelphia, PA, USA. bailisw@email.chop.edu.

PMID: 31776929
DOI: 10.1007/978-1-0716-0203-4_14

Abstract

CRISPR-Cas9 technology allows for rapid, targeted genome editing at nearly any loci with limited off-target effects. Here, we describe a method for using retroviral transduction to deliver single-guide RNA to primary bone marrow-derived macrophages. This protocol allows for high-throughput reverse genetics assays in primary immune cells and is also compatible with retroviral systems for transgene expression.

Keywords: Bone marrow; CRISPR/Cas9; Gene targeting; Macrophage; Mice; Retrovirus.

MeSH terms

Animals
CRISPR-Associated Protein 9 / metabolism*
CRISPR-Cas Systems / genetics*
Cells, Cultured
Gene Targeting / methods*
HEK293 Cells
Humans
Macrophages / metabolism*
Mice
Retroviridae / metabolism
Transduction, Genetic

Substances

CRISPR-Associated Protein 9