2.4. Generation of CRISPR/Cas9-Mediated Knockout (KO) Cell Line

CRISPR/Cas9-mediated gene editing was performed using a px459 vector (Addgene, Catalog No. 48139) targeting murine TREM2 or NLRP3 in BV2 cells, and the plasmid construction has been previously described [24]. The sgRNAs targeting the TREM2 sequence (5′-TCCCAAGCCCTCAACACCA-3′) and NLRP3 sequence (5′-CAAGCTGGCTCAGTATCTAG-3′) were synthesized by Comate Bioscience Company (Shanghai, China). According to the manufacturer’s instructions, plasmids were transfected with Attractene Transfection Reagent (QIAGEN, Beijing, China, Catalog No. 301004). Clonal lines were established by 96-well plate screening. The KO cell lines were verified by western blotting after the clones had formed. For the high glucose treatment experiment, KO cells were treated with high glucose at 35 mM for 12 h.