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Author Spotlight: Efficient CRISPR/Cas9 Genome Editing in Bone Marrow-Derived Macrophages for Precise Gene Disruption
Published on: August 4, 2023
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CRISPR/Cas9 Gene Targeting in Primary Mouse Bone Marrow-Derived Macrophages
1Division for Protective Immunity, Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia, Philadelphia, PA, USA. bailisw@email.chop.edu.
Methods in Molecular Biology (Clifton, N.J.)
|November 29, 2019
Summary
CRISPR-Cas9 gene editing enables precise DNA modification. This study details a method to deliver CRISPR components into immune cells, facilitating genetic studies and transgene expression.
Area of Science:
- Molecular Biology
- Genetics
- Immunology
Background:
- CRISPR-Cas9 is a powerful tool for targeted genome editing.
- Primary immune cells are crucial for biological research but challenging to genetically manipulate.
- Efficient delivery of gene editing components is essential for functional studies.
Purpose of the Study:
- To develop a method for delivering CRISPR-Cas9 components into primary bone marrow-derived macrophages.
- To enable high-throughput reverse genetics assays in primary immune cells.
- To ensure compatibility with existing retroviral systems for transgene expression.
Main Methods:
- Utilizing retroviral transduction to deliver single-guide RNA (sgRNA).
- Targeting primary bone marrow-derived macrophages.
- Implementing CRISPR-Cas9 technology for genome editing.
Main Results:
- Successful delivery of sgRNA to primary macrophages.
- Demonstration of CRISPR-Cas9's capability in primary immune cells.
- Compatibility with retroviral transgene expression systems.
Conclusions:
- The described method provides an efficient way to perform CRISPR-Cas9 gene editing in primary immune cells.
- This protocol supports high-throughput reverse genetics in macrophages.
- The approach is versatile and integrates with established retroviral techniques for broader applications.
