An inducible CRISPR (clustered regulatory interspaced short palindrome repeats) /Cas9 (CRISPR-associated 9) lentiviral system was used to construct THP-1 ZBTB16 mutant lines. THP-1 cells were infected by lentiviral particles containing the pFUCas9-mCherry vector and sgRNAs targeting exon 2 of ZBTB16, cloned into the doxycycline (DOX)-inducible vector pFgh1tUTG-GFP. THP-1 cells expressing Cas9 and sgRNA were isolated as GFP and mCherry positive cells by flow cytometry (FACS Aria II, BD Biosciences)61. Gene knockout was induced by the culture of transduced cells with 1 μg/mL DOX (Sigma) for 5 days and confirmed by measuring ZBTB16 mRNA levels by RT-PCR (sgRNA and RT-PCR primer sequences are shown in Supplementary Table S1).
