Optimized protocol for CRISPR knockout of human iPSC-derived macrophages.
STAR Protoc
; 5(1): 102903, 2024 Mar 15.
Article
in En
| MEDLINE
| ID: mdl-38401123
ABSTRACT
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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Neurodegenerative Diseases
/
Induced Pluripotent Stem Cells
Limits:
Humans
Language:
En
Journal:
STAR Protoc
Year:
2024
Document type:
Article