CRISPR/Cas9-Mediated Genome Engineering of Primary Human B Cells

Kanut Laoharawee; Matthew J Johnson; Branden S Moriarity

doi:10.1007/978-1-0716-0290-4_24

CRISPR/Cas9-Mediated Genome Engineering of Primary Human B Cells

Methods Mol Biol. 2020:2115:435-444. doi: 10.1007/978-1-0716-0290-4_24.

Authors

Kanut Laoharawee¹, Matthew J Johnson¹, Branden S Moriarity²

Affiliations

¹ Department of Pediatrics, Cardiovascular Research, Masonic Cancer Center, Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA.
² Department of Pediatrics, Cardiovascular Research, Masonic Cancer Center, Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA. mori0164@umn.edu.

PMID: 32006415
DOI: 10.1007/978-1-0716-0290-4_24

Abstract

The CRISPR/Cas9 system allows for site-specific gene editing and genome engineering of primary human cells. Here we describe methods for gene editing and genome engineering of B cells isolated from human peripheral blood mononuclear cells using CRISPR/Cas9. Editing frequencies of up to 90% and integration rates greater than 60% can be achieved with this method.

Keywords: B-cell engineering; CRISPR/Cas9; Genome engineering; Primary human B cells; rAAV.

MeSH terms

B-Lymphocytes / cytology
B-Lymphocytes / metabolism*
CRISPR-Cas Systems*
Cells, Cultured
Clustered Regularly Interspaced Short Palindromic Repeats
Gene Editing / methods*
Humans
Leukocytes, Mononuclear / cytology
Leukocytes, Mononuclear / metabolism