| Products/Services Used | Details | Operation |
|---|---|---|
| Plasmid DNA Preparation | All-in-one CRISPR/Cas9-gRNA plasmids (pLentiCRISPR-v2) were purchased from GenScript. Plasmids were extracted using Qiagen Plasmid Maxi Kit. HEK293T cells were transfected with the plasmids to package lentiviruses using TransIT®- LT1 Transfection Reagent (Mirus Bio) according to the manufacturer's instructions. The collected lentiviruses were used to infect organoid cultures to silence genes of interest. Puromycin (2 μg/mL, Thermo Fisher Scientific) was added to the cell culture medium for selection. | Get A Quote |
Colorectal cancer is a leading cause of cancer deaths. Most colorectal cancer patients eventually develop chemoresistance to the current standard-of-care therapies. Here, we used patient-derived colorectal cancer organoids to demonstrate that resistant tumor cells undergo significant chromatin changes in response to oxaliplatin treatment. Integrated transcriptomic and chromatin accessibility analyses using ATAC-Seq and RNA-Seq identified a group of genes associated with significantly increased chromatin accessibility and upregulated gene expression. CRISPR/Cas9 silencing of fibroblast growth factor receptor 1 (FGFR1) and oxytocin receptor (OXTR) helped overcome oxaliplatin resistance. Similarly, treatment with ... More
