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Related Concept Videos

CRISPR01:59

CRISPR

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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CRISPR/Cas9 Genome Editing01:28

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Updated: Dec 29, 2025

Genome Engineering of Primary Human B Cells Using CRISPR/Cas9
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Gene Editing in B-Lymphoma Cell Lines Using CRISPR/Cas9 Technology.

Baoyan Bai1,2, June Helen Myklebust3,4, Sébastien Wälchli5

  • 1Department of Cancer Immunology, Institute for Cancer Research, Oslo University Hospital, Oslo, Norway. baoyan.bai@rr-research.no.

Methods in Molecular Biology (Clifton, N.J.)
|February 2, 2020
PubMed
Summary

This study introduces a sequential delivery method for CRISPR/Cas9 components, enhancing genome editing efficiency in challenging suspension cells. The strategy improves on-target editing and minimizes off-target effects for broader CRISPR applications.

Keywords:
B-lymphoma cellsCRISPR/Cas9Cas9-expressing cellsGenome editingSquare wave electroporation

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR/Cas9 technology has revolutionized genome editing in eukaryotes.
  • Efficient delivery of CRISPR components into suspension cell lines, such as B-lymphoid cells, remains a significant challenge.
  • Existing methods often struggle with low efficiency and potential off-target mutations.

Purpose of the Study:

  • To develop an improved method for CRISPR/Cas9 delivery into difficult-to-edit suspension cells.
  • To enhance the efficiency and specificity of genome editing in B-lymphoid cell lines.
  • To provide a versatile strategy applicable to various cell types.

Main Methods:

  • Sequential delivery of Cas9 and single-guide RNA (sgRNA) components.
  • Stable Cas9 expression achieved via retroviral transduction.
  • Transient delivery of sgRNA into Cas9-expressing cells.

Main Results:

  • Improved on-target genome editing efficiency in B-lymphoid cells.
  • Reduced off-target mutations due to the transient nature of sgRNA presence.
  • Demonstrated applicability to cell types challenging for CRISPR/Cas9 editing.

Conclusions:

  • The sequential delivery strategy effectively overcomes challenges in editing suspension cells.
  • This method enhances CRISPR/Cas9 precision and efficiency.
  • The approach offers a valuable tool for broader genome engineering applications.