RNA-seq of intestinal crypts vs intestinal smooth muscle with a specific MMP17 deletion compared to wild-type

We wanted to assess the role of a specific smooth muscle protein (MMP17) in two different intestinal compartments, the epithelium (crypts) and the smooth muscle. To do that we isolate intestinal crypts from wild-type (WT) and knockout (KO, Mmp17-/-) mice, and obtained clean strips of smooth muscle. After muscle dissociation, we obtained RNA directly from crypts and muscle, and it was used for RNA-seq. By comparing WT and KO samples we observed a higher impact in gene expression affecting crypts, even though MMP17 is only expressed in muscle. This helped us to identify altered signaling pathways in KO crypts that linked MMP17 with SMAD4 and BMP signaling.本研究旨在探究特定平滑肌蛋白（MMP17）在肠道两大分区——上皮组织（隐窝，crypts）和平滑肌——中的作用。为此，我们分别从野生型（wild-type, WT）及基因敲除型（knockout, KO, Mmp17-/-）小鼠体内分离肠道隐窝，并获取纯净的平滑肌条带。完成平滑肌解离操作后，我们直接从隐窝及平滑肌组织中提取总RNA，将其用于RNA测序（RNA-seq）实验。通过对比野生型与基因敲除型样本的测序结果，我们发现尽管MMP17仅在平滑肌组织中表达，但其缺失对隐窝的基因表达产生了更为显著的影响。该发现帮助我们明确了基因敲除型小鼠隐窝中发生异常的信号通路，揭示了MMP17与SMAD4及骨形态发生蛋白（bone morphogenetic protein, BMP）信号通路之间的关联。