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Genome Engineering of Primary Human B Cells Using CRISPR/Cas9

Article DOI: 10.3791/61855-v • 08:20 min • November 3rd, 2020
November 3rd, 2020 •
1,2,3, 1,2,3, 1,2,3, 1,2,3, 1,2,3, 1,2,3
1Department of Pediatrics, University of Minnesota, 2Center for Genomic Engineering, University of Minnesota, 3Masonic Cancer Center, University of Minnesota

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Here we provide a detailed, step-by-step protocol for CRISPR/Cas9-based genome engineering of primary human B cells for gene knockout (KO) and knock-in (KI) to study biological functions of genes in B cells and the development of B-cell therapeutics.

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Genome Engineering Primary Human B Cells CRISPR/Cas9 Gene Elimination Loss Of Function Effects Transgene Expression Research Therapeutic Purposes Universal Platform High Efficiencies Recombinant Antibodies Enzymes Infections Enzymopathies Thawing B Cells FBS Sterile Conical Tube Incubation Resuspension Sterile PBS Centrifugation Cell Pellet B-cell Expansion Medium
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