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

DOI :

10.3791/62328-v

November 13th, 2021

November 13th, 2021

8,908 Views

1The Laboratory of Genetic Regulators in the Immune System, School of Laboratory Medicine, Xinxiang Medical University, 2Centre d’Immunologie de Marseille-Luminy, Aix Marseille Université

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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