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Here, we present a protocol for lentiviral delivery of CRISPR-Cas9 to human induced pluripotent stem cell (iPSC)-derived macrophages using co-incubation with VPX virus-like particles (VPX-VLPs). We describe steps for producing polybrene and puromycin kill curves, VPX viral production, and VPX-VLP titration by western blotting. We then detail procedures for iPSC macrophage precursor lentiviral transduction and lentiviral CRISPR-Cas9-based knockout in iPSC-derived macrophages. This protocol uses efficient genome-editing techniques to explore macrophage involvement in immune response, chronic inflammation, neurodegenerative disease, and cancer progression. For complete details on the use and execution of this protocol, please refer to Navarro-Guerrero et al.1.

Original publication

DOI

10.1016/j.xpro.2024.102903

Type

Journal article

Journal

STAR Protoc

Publication Date

15/03/2024

Volume

5

Keywords

CRISPR, High Throughput Screening, Microbiology, Molecular Biology, Humans, CRISPR-Cas Systems, Induced Pluripotent Stem Cells, Neurodegenerative Diseases, Gene Editing, Macrophages