Elf4−/− HEK293 cells, Hipk2−/− MEF cells, and Hipk2−/−, Irf3−/−, Irf7−/−, p65−/− iBMDMs were generated by CRISPR-Cas9 knockout. Specific guide RNAs were ligated into the BsmB1 restriction site of the inducible lentiviral vector (lentiGuide-Puro). Lentivirus particles were produced by cotransfected HEK293 cells with guide RNA plasmids (2 μg), packaging plasmids pVSVg (800 ng, AddGene 8454), and psPAX2 (800 ng, AddGene 12260). At 24 hours after transfection, the medium was changed, and viral supernatant was collected at 48 to 72 hours. A total of 1 × 105 iBMDMs or MEF cells were infected with 2 ml of viral supernatant supplemented with polybrene (8 μg/ml) and incubated for 48 hours. Knockout cells were selected by puromycin (1 μg), and each monoclone was confirmed by sequencing. Cells were negative for mycoplasma. The specific guide RNA sequences are listed in table S1.

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