Project description:Our RNAseq results showed that P2RY6 induced the expression of Ptgs1 and Ptgs2 in MC38 cells. To confirm whether P2RY6 increases the opening of cis-regulatory elements to promote Ptgs1/2 transcription, we performed Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq) to determain the chromatin accessibility across the genome in MC38 cells with or without P2RY6 expression.
Project description:To investigate the effect of different tumor-associated glycans, mouse colorectal cancer cells (MC38 cell line) were genetically modified using CRISPR-Cas9 and CRISPR-dCas-VPR technology targeting different genes involved in glycosylation pathways. To study the effect of altered glycosylation at the transcriptional level, next generation sequencing was performed on the entire panel of glycovariant MC38 cell lines. The MC38 knockout cell lines created with the CRISPR-Cas9 technology are: MC38-MOCK#1, MC38-CMAS KO, MC38-COSMC KO, MC38-CMAS+COSMC KO. The MC38 overexpressing cell lines generated with the CRISPR-dCas9-VPR technology are: MC38-MOCK#2, MC38-FUT4, MC38-FUT9. Details on the cell lines can be found in the corresponding manuscript(s).
Project description:We investigate the single-cell landscape of the inflammatory mouse tumor model MC38, a C57BL/6 tumor cell line derived from colon adenocarcinoma. MC38 (diluted in HBSS and matrigel) was inoculated in the right unilateral flank (in the border of positions B2 and B3) of C57BL/6 mice (ref Study 16-3384 AV). Draining lymph nodes were taken one day after group-out (average 150-250 mm3 tumor size at day 0), approximately 14-19 days. Draining lymph nodes were dissociated and flow sorted accordingly to obtain the cells for 10x Chromium 5' Gene Expression Profiling and TCR sequencing.
Project description:We investigate the single-cell landscape of the inflammatory mouse tumor model MC38, a C57BL/6 tumor cell line derived from colon adenocarcinoma. MC38 (diluted in HBSS and matrigel) was inoculated in the right unilateral flank (in the border of positions B2 and B3) of C57BL/6 mice (ref Study 16-3384 AV). Draining lymph nodes were taken one day after group-out (average 150-250 mm3 tumor size at day 0), approximately 14-19 days. Draining lymph nodes were dissociated and flow sorted accordingly to obtain the cells for 10x Chromium 5' Gene Expression Profiling.
Project description:We designed a novel intratumoral (IT) IL-12 mRNA therapy to promote local IL-12 tumor production whilst mitigating systemic effects. A single IT dose of mouse (m)IL-12 mRNA induced IFNγ and CD8+ T cell-dependent tumor regression in multiple syngeneic mouse models including MC38-R, and animals with a complete response demonstrated immunity to re-challenge. In order to further investigate the impact of mIL-12 mRNA on the MC38-R TME, we performed tumor transcriptomic evaluation 24h and 7 days after a single IT dose of 0.5 micrograms IL-12 or control mRNA. mRNA was isolated from snap-frozen syngeneic tumors using an RNeasy mini kit (Qiagen) and quantified on the Affymetrix mouse 430 2.0 microarray.
