Multiome snRNA + snATAC on MCF10-2A cells to study how high CENP-A levels promote EMT
CENP-A, the centromeric histone H3 variant, is essential for the proper segregation of human chromosomes to daughter cells. High CENP-A expression leads to the misincorporation of CENP-A in non-centromeric chromatin and can promote an epithelial-mesenchymal transition (EMT).  Here, we explored how CENP-A could promote EMT, considering both its impact on transcription and chromatin structure. For this, we used a MCF10-2A TetOn-CENPA-FLAG-HA cell line where high CENP-A expression can be induced by Doxycycline (Dox) treatment. This cell line is p53-defective (transduction with a dominant-negative (DN) p53 vector). Upon CENP-A overexpression, these cells progressively accumulate mesenchymal states. For the same nuclei, we jointly profiled single-nucleus RNA (snRNA) and single-nucleus ATAC (snATAC) data using 10X Multiome at days 10 and 24 (prior and after the mesenchymal states accumulation). In order to profile changes in transcription and cell states, we compared cells expressing either basal CENP-A levels (non-induced, control with no Dox), or high CENP-A levels (induced with 10 ng/ml Dox).
CENP-A, the centromeric histone H3 variant, is essential for the proper segregation of human chromosomes to daughter cells. High CENP-A expression leads to the misincorporation of CENP-A in non-centromeric chromatin and can promote an epithelial-mesenchymal transition (EMT).  Here, we explored how CENP-A could promote EMT, considering both its impact on transcription and chromatin structure. For this, we used a MCF10-2A TetOn-CENPA-FLAG-HA cell line where high CENP-A expression can be induced by Doxycycline (Dox) treatment. This cell line is p53-defective (transduction with a dominant-negative (DN) p53 vector). Upon CENP-A overexpression, these cells progressively accumulate mesenchymal states. For the same nuclei, we jointly profiled single-nucleus RNA (snRNA) and single-nucleus ATAC (snATAC) data using 10X Multiome at days 10 and 24 (prior and after the mesenchymal states accumulation). In order to profile changes in transcription and cell states, we compared cells expressing either basal CENP-A levels (non-induced, control with no Dox), or high CENP-A levels (induced with 10 ng/ml Dox).
European Bioinformatics Institute