METTL14 regulates chondrogenesis through the GDF5-RUNX-extracellular matrix gene axis during limb development [scRNA-seq]

Mettl14 forms a protein complex that induces m6A methylation on RNA but its roles in mouse limb development remains elusive. Here, the Mettl14 gene was knocked out in a limb bud-specific manner in Mettl14 flox/flox mice with the Prx1 promoter-driven cre. Dissociated cells from day 12.5 (E12.5) embryonic limb buds were applied to scRNA-seq to identify differentially expressed genes in a cell type-specific manner. Samples prepared from Per1cre;Mettl14 wild type/flox and Per1cre;Mettl14 flox/flox mice were labeled as cHet and cKO, respectively. Single cells prepared from limb buds on E12.5 were applied to scRNA-seq.Mettl14可形成诱导RNA发生m6A甲基化（m6A methylation）的蛋白质复合物，但其在小鼠肢体发育中的功能仍尚不明确。本研究通过Prx1启动子驱动的Cre重组酶（Cre recombinase），在Mettl14 flox/flox小鼠中以肢体芽特异性的方式敲除Mettl14基因。取胚胎发育第12.5天（E12.5）的胚胎肢体芽解离细胞进行单细胞RNA测序（scRNA-seq），以细胞类型特异性的方式鉴定差异表达基因。由Per1cre;Mettl14野生型/flox和Per1cre;Mettl14 flox/flox小鼠制备的样本分别被标记为条件性杂合（cHet）和条件性敲除（cKO）。从E12.5胚胎肢体芽制备的单细胞被用于单细胞RNA测序（scRNA-seq）。