title: Chromatin architecture profiling of a model of CRISPR-mediated insulator disruption in the MMP locus in triple-negative breast cancer cell lines
design: Chromatin architecture profiling of a model of CRISPR-mediated insulator disruption in the MMP locus in triple-negative breast cancer cell lines
library_construction_protocol: 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.