Gene expression in murine cartilage, 3 months of age, wild type vs miR-455 null

MicroRNA-455 is a key microRNA in cartilage, strongly induced by Sox9. Using CrispR-Cas9, we produced a global deletion of miR-455 in the mouse and characterised cartilage and bone. In this study, we compared the cartilage gene expression in 3 month old mice, between wild-type and miR-455 null. Cartilage was dissected under a microscope from both tibia and femur of the hind limb into RNALater, snap frozen in liquid nitrogen and stored at -80C. Cartilage from two mouse knees per sample was pooled and ground to a fine powder under liquid nitrogen; RNA was purified using a combination of Trizol and miRVana We then performed RNASeq analysis on tissue from n=3 wild-type vs miR-455 mice微小RNA-455（microRNA-455）是软骨中的关键微小RNA，可被Sox9强力诱导表达。本研究通过CRISPR-Cas9技术在小鼠体内构建miR-455全局敲除模型，并对其软骨与骨骼组织进行表征分析。本实验比较了3月龄野生型与miR-455敲除型小鼠的软骨基因表达差异。具体样本处理流程如下：从后肢胫骨与股骨处显微分离软骨，将其转移至RNALater保存液中，经液氮快速冷冻后储存于-80℃环境。每份样本混合2只小鼠的膝关节软骨，在液氮条件下研磨为细粉；采用Trizol与miRVana试剂盒联合提取总RNA。随后对n=3例野生型小鼠与n=3例miR-455敲除型小鼠的组织样本开展RNA测序（RNA-seq）分析。