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Gene Knock-in by CRISPR/Cas9 and Cell Sorting in Macrophage and T Cell Lines
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Gene Knock-in by CRISPR/Cas9 and Cell Sorting in Macrophage and T Cell Lines

Gene Knock-in by CRISPR/Cas9 and Cell Sorting in Macrophage and T Cell Lines

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11:32 min

November 13, 2021

DOI:

11:32 min
November 13, 2021

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Summary

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This protocol uses fluorescent reporters and cell sorting to simplify knock-in experiments in macrophage and T cell lines. Two plasmids are used for these simplified knock-in experiments, namely a CRISPR/Cas9- and DsRed2-expressing plasmid and a homologous recombination donor plasmid expressing EBFP2, which is permanently integrated at the Rosa26 locus in immune cells.

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