Project description:MCF10A cells were CRISPR-Cas9 edited to create heterozygous deletion in RAD21 and SMC3 subunits of cohesin. STAG2 is on the X chromosome, hence CRISPR-Cas9 editing resulted in complete loss of STAG2. Total RNA was sequenced from the MCF10A parental and cohesin mutant MCF10A lines. The acute megakaryoblastic leukaemia cell line CMK was CRISPR-Cas9 edited to cotain STAG2 R614* mutation. CRISPR-Cas9 edited STAG2 mutant line showed complete loss of STAG2. CMK parental and the STAG2 mutant line were treated with Wnt3a for 4 hours and total RNA was sequenced at in the control or non-treated (con) and following 4 hours of Wnt3a treatment (Wnt3a4hr).
Project description:We performed a CRISPR-based functional genetic screen targeting TEAD4 binding motif located within the YAP-bound enhancers. The screen identified seven functional enhancers whose targeting resulted in a marked negative effect on the proliferation of MCF10A-YAP-5SA cells (overexpressed constrictively activated YAP) but no significant effect on MCF10A (parental control) cells. We then carried out RNA-seq analysis on MCF10A-YAP-5SA cells transduced with sgRNA vectors targeting these seven enhancers (enhancers A, B, C, D, E, F, G) as well as the non-targeting sgRNA as control (NT).
Project description:The overall goal of this study is to identify the genomic binding of RUNX1 in MCF10A cells. We used ChIPseq (chromatin immunoprecipitation assay followed by deep sequencing) to identify the binding sites of RUNX1 in MCF10A cells. We performed ChIPseq of RUNX1 using parental MCF10A cells and did not identify high confident binding sites. To overcome this hurdle, we first generated a RUNX1 deleted MCF10A cell line using CRISPR-Cas9. We then transduced this RUNX1 KO MCF10A cells with lentiviruses that inducibly expresses RUNX1. After treating RUNX1 inducible MCF10A cells with 1 ug/ml doxycycline for 24 hours, we performed ChIPseq of RUNX1.
Project description:This dataset contains ChIP-seq data profiling genomic binding of H3K27ac and H3K4me3 in single cell-derived control, as well as CRISPR/Cas9 induced tRNA gene deletion clones and intergenic region deletion clones in human cancer cell lines HAP1. In this study, we found a large genomic deletion of 10q23 in Cas9 modified clones and further investigate the effect of H3K27ac binding.
Project description:We performed a CRISPR-based functional genetic screen targeting TEAD4 binding motif located within the YAP-bound enhancers. The screen identified a novel enhancer regulating TRAM2. We observed TRAM2 overexpression can induce EMT as YAP does. Then, we performed the RNA-seq to measure the transcription changes upon TRAM2-KOs in MCF10A-YAP cells
Project description:This dataset contains ploy-A tailed enriched RNA-seq data obtained from single cell-derived control and CRISPR/Cas9 induced tRNA gene deletion clones in the human cancer cell line HAP1. In this study, we found a large genomic deletion of the 10q23 locus in our Cas9 modified clones and further investigate the effect on the transcriptome.
