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Genome Engineering of Primary Human B Cells Using CRISPR/Cas9
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JoVE Journal Biologia
Genome Engineering of Primary Human B Cells Using CRISPR/Cas9

Genome Engineering of Primary Human B Cells Using CRISPR/Cas9

DOI:
10.3791/61855-v

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08:20 min

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November 03, 2020

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, , , , ,
1Department of Pediatrics,University of Minnesota, 2Center for Genomic Engineering,University of Minnesota, 3Masonic Cancer Center,University of Minnesota

Capitoli

  • 00:04Introduction
  • 00:56Human B-cell Expansion
  • 02:25Primary Human B-cell Engineering
  • 05:30Results: CRISPR/Cas9-based Genome Engineering of Primary Human B Cells for Gene Knockout and Knock-in
  • 07:33Conclusion

Summary

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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.

Tags

Genome EngineeringCRISPR/Cas9Primary Human B CellsGene KnockoutGene InsertionRecombinant AntibodyEnzymopathyCell CultureElectroporationTransfectionSgRNACas9 MRNA

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