EXPERIMENT_TITLE: Chromatin architecture profiling of a model of CRISPR-mediated insulator disruption in the MMP locus in triple-negative breast cancer cell lines
EXPERIMENT_DESIGN_LIBRARY_DESCRIPTOR_LIBRARY_CONSTRUCTION_PROTOC: Cells were collected using 0.25% Trypsin-EDTA 1x (Gibco) Cells were cultured using RPMI 1640 media complemented with 10% FBS and penicillin/streptomycin at 5% CO2 37ºC. Cells were transiently transfected with either empty pSpCas9 or pSpCas9 containing a sgRNA sequence against insulator element 8. Transfection was performed using Lipofectamine3000 following manufacturer's guidelines. Cells were then sorted using FACSAria Fusion using GFP+ as a reporter. The established models were then subcultured to perform high throughput experiments. DNA was extracted following the manufacturer's protocol from Dovetail Genomics, now Cantata Bio. Library prep was done using the NEBNext Ultra II DNA Library Prep kit for Illumina.
STUDY_DESCRIPTOR_STUDY_ABSTRACT: Using CRISPR-Cas9 technology, we stably disrupted an insulator element in two different TNBC cell lines. The aim of this experiment was to characterize the chromatin architecture profile using Hi-C. For this experiment, we selected three biological replicates of MDA-MB-231 (WT and IE8dis).
SAMPLE_TITLE: 436-IE8dis-1
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