| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | HCT 116 stably expressing Cas9-GFP were obtained by lentiviral transduction with pLentiCRISPR-EF1αs-Cas9-P2A-GFP-PGK-puro. KBM-7 infected with dox-inducible Cas9 were obtained by sequential retroviral transduction with pWPXLd-EF1A-rtTA3-IRES-EcoRec-PGK-Puro and pSIN-TRE3G-Cas9-P2A-GFP PGK-Blast. pLVX-3xFLAG-STAG1r-IRES-Puro and pLVX-3xFLAG-STAG2r-IRES-Puro lentiviral vectors for siRNA-resistant transgene expression were generated by gene synthesis (GenScript) based on the STAG1 cDNA sequence NCBI NM_005862.2 and STAG2 cDNA sequence NCBI NM_001042749.2 followed by cloning into the parental pLVX vectors (Clontech). Silent nucleotide changes were introduced into the STAG1 and STAG2 coding sequences within the siRNA target sequences to render the transgenes siRNA-resistant. For competition assays, U6-sgRNA-EF1αs-mCherry-P2A-neo lentiviral vectors were used. | Get A Quote |
Recent genome analyses have identified recurrent mutations in the cohesin complex in a wide range of human cancers. Here we demonstrate that the most frequently mutated subunit of the cohesin complex, STAG2, displays a strong synthetic lethal interaction with its paralog STAG1. Mechanistically, STAG1 loss abrogates sister chromatid cohesion in STAG2 mutated but not in wild-type cells leading to mitotic catastrophe, defective cell division and apoptosis. STAG1 inactivation inhibits the proliferation of STAG2 mutated but not wild-type bladder cancer and Ewing sarcoma cell lines. Restoration of STAG2 expression in a mutated bladder cancer model alleviates the dependency on STAG1. Thus, STAG1 and STAG2 act redu... More
