RNA-seq analysis of Mst1/2 deleted bronchiolar epithelial cells from adult mouse lungs. Mus musculus

Mst1 and Mst2 were conditionally deleted from non-ciliated bronchiolar epithelial cells in the mature lung. Bronchiolar epithelial cells from control and Mst1/2 deleted mice were isolated by cell sorting and used for RNA-seq analysis. Overall design: Scgb1a1-rtTA/tetO-Cre/Mst1;2-flx/flx (Mst1/2 D/D) mice were generated to conditionally delete Mst1 and Mst2 from non-ciliated, secretory bronchiolar epithelial cells. Adult mice were maintained on doxycycline food for 16 days to induce deletion of Mst1/2. Lin-/CD326+/CD24-intermediate cells were isolated by fluorescence cell sorting to enrich for the targeted airway epithelial cells. mRNA isolated from Lin-/CD326+/CD24-intermediate cells from control and Mst1/2 D/D mice was pooled and analyzed by RNA-seq to identify transcriptional changes following deletion of Mst1 and Mst2 from mature lung bronchiolar epithelial cells.

本研究在成熟肺脏的非纤毛细支气管上皮细胞中条件性敲除了Mst1与Mst2基因。通过细胞分选技术分离对照组与Mst1/2敲除小鼠的细支气管上皮细胞，用于RNA测序（RNA-seq）分析。总体实验设计：构建Scgb1a1-rtTA/tetO-Cre/Mst1;2-flx/flx（Mst1/2 D/D）小鼠，以实现从非纤毛分泌型细支气管上皮细胞中条件性敲除Mst1与Mst2。将成年小鼠饲喂含多西环素的饲料16天，以诱导Mst1/2基因敲除。通过荧光细胞分选分离Lin-/CD326+/CD24中间型细胞，以富集目标气道上皮细胞。将对照组与Mst1/2 D/D小鼠的Lin-/CD326+/CD24中间型细胞提取的mRNA混合后，采用RNA-seq进行分析，以鉴定成熟肺脏细支气管上皮细胞中Mst1与Mst2敲除后发生的转录组变化。