Project description:The airway mucociliary epithelium is consituted of three main cell types : columnar ciliated plus secretory cells and basal cells. Columnar cells are represented by a great majority of ciliated cells. We used Cell sorting by FACSaria to separate basal cells from ciliated and secreting columnar cells. Then, we performed microRNA high throughput sequencing to investigate the specific signature of microRNA of basal and columnar cells. Overall design: miRNAs high throughput sequencing profiling of human nasal mucosa: basals cells (B) and columnars (C) cells for 3 donors.
Project description:High-throughput genome-wide translocation sequencing (HTGTS) is a robust approach to identify genome-wide translocation junctions. We performed HTGTS to study the fate of introduced c-myc DSBs in mouse splenic B cells activated for activation cytidine deaminase (AID)-dependent class switch recombination (CSR). We found frequent translocations of c-myc DSBs to AID-initiated DSBs in IgH switch regions in wild-type (WT) and ATM-deficient B cells. However, c-myc also translocated frequently to newly generated DSBs within a 35-megabase region downstream of IgH in ATM-deficient, but not WT, CSR-activated B cells. Moreover, we found such DSBs and translocations in activated B cells that did not express AID or undergo CSR. These findings indicate that ATM deficiency leads to formation of chromosome 12 dicentrics via RAG-initiated IgH DSBs in progenitor B cells and that these dicentrics can be propagated developmentally into mature B cells where they generate new DSBs downstream of IgH via breakage-fusion-bridge cycles. Preparation of libraries from WT or ATM-deficient activated by a-CD40/IL4 or RP105.