SIM2s-FLAG SUM159 and shSIM2 MCF7 cells were generated as previously described36 (link),44 (link). The CRISPR/Cas9 SIM2 KO vector (pLV[2CRISPR]-hCas9:T2A:Puro-U6 > hSIM2[gRNA#1]-U6 > hSIM2[gRNA#2]) was designed with two gRNA sequences targeting exon 1 in hSIM2 (gRNA#1 5’-TCCGCGCTACTTCCTCTTCA-3’; gRNA#2 5’-ATGCGCGCCGTCTTCCCCGA-3’) and was generated and transformed into Stbl3 competent cells by VectorBuilder.
Plasmid DNA was isolated using a ZymoPURE Plasmid DNA Isolation Kit (Zymo Research). Lentiviral transduction was performed as previously described, and cells were selected with 2 μg/mL puromycin (Sigma‒Aldrich) for 7 days or after five passages.
Plasmid DNA was isolated using a ZymoPURE Plasmid DNA Isolation Kit (Zymo Research). Lentiviral transduction was performed as previously described, and cells were selected with 2 μg/mL puromycin (Sigma‒Aldrich) for 7 days or after five passages.
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