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Gene Editing in B-Lymphoma Cell Lines Using CRISPR/Cas9 Technology

  • Baoyan BaiEmail author
  • June Helen Myklebust
  • Sébastien Wälchli
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2115)

Abstract

Genome editing in eukaryotes has greatly improved through the application of targeted editing tools. The development of the CRISPR/Cas9 technology has facilitated genome editing in mammalian cells. However, efficient delivery of CRISPR components into cells growing in suspension remains a challenge. Here, we present a strategy for sequential delivery of the two essential components, Cas9 and sgRNA, into B-lymphoid cell lines. Stable Cas9 expression is obtained by retroviral transduction, before sgRNA is transiently delivered into the Cas9+ cells. This method improves the on-target efficiency of genome editing and, through the transient presence of sgRNA, reduces the potential off-target sites. The current method can be easily applied to other cell types that are difficult to edit with CRISPR/Cas9.

Key words

B-lymphoma cells CRISPR/Cas9 Genome editing Cas9-expressing cells Square wave electroporation 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Baoyan Bai
    • 1
    • 2
    Email author
  • June Helen Myklebust
    • 1
    • 2
  • Sébastien Wälchli
    • 3
  1. 1.Department of Cancer Immunology, Institute for Cancer ResearchOslo University HospitalOsloNorway
  2. 2.KG. Jebsen Centre for B cell MalignanciesUniversity of OsloOsloNorway
  3. 3.Department of Cellular Therapy, Division of Cancer MedicineOslo University HospitalOsloNorway

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