Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China.
Genes (Basel). 2022 Apr 9;13(4):663. doi: 10.3390/genes13040663.
Myogenesis is a complex process controlled by several coding and non-coding RNAs (ncRNAs), such as circular RNAs (circRNAs) that are known to function as endogenous microRNAs (miRNAs) sponges. Cerebellar Degeneration-Related protein 1 antisense (CDR1as) is the most spotlighted circRNA that is known as an miR-7 sponge, which has bloomed circRNAs' research in animal disease and physiology. Here, we screened for miRNAs and mRNA associated with CDR1as and further characterized their regulatory function during muscle differentiation. We found that a total of 43 miRNAs (including miR-107-3p, miR-125b-5p, miR-140-5p, miR-29a-3p, and miR-27a-3p upregulated) and 789 mRNAs (including ANGPT1, E2F2, CCN1, FGFR1, and MEF2C downregulated) were differentially expressed in goat skeletal muscle satellite cells (SMSCs). Further, knockdown of CDR1as and ANGPT1 inhibited SMSCs differentiation. miR-27a-3p was differentially upregulated after the knockdown of CDR1as in SMSCs. Overexpressed miR-27a-3p decreased SMSCs differentiation. Via RNAhybrid and luciferase, miR-27a-3p was identified to regulate ANGPT1. We discovered that miR-27a-3p has an inverse relationship with CDR1as and decreases the expression level of ANGPT1 during SMSCs differentiation. In summary, our study demonstrates that siCDR1as inhibits myoblast differentiation by downregulating ANGPT1 mRNA via miR-27a-3p in SMSCs.
肌发生是一个由几种编码和非编码 RNA(ncRNA)控制的复杂过程,例如已知作为内源性 microRNA(miRNA)海绵的环状 RNA(circRNA)。小脑退化相关蛋白 1 反义(CDR1as)是最受关注的 circRNA,作为 miR-7 的海绵,它促进了动物疾病和生理学中 circRNAs 的研究。在这里,我们筛选了与 CDR1as 相关的 miRNA 和 mRNA,并进一步表征了它们在肌肉分化过程中的调节功能。我们发现,在山羊骨骼肌卫星细胞(SMSCs)中,共有 43 个 miRNA(包括 miR-107-3p、miR-125b-5p、miR-140-5p、miR-29a-3p 和 miR-27a-3p 上调)和 789 个 mRNA(包括 ANGPT1、E2F2、CCN1、FGFR1 和 MEF2C 下调)差异表达。进一步,CDR1as 和 ANGPT1 的敲低抑制了 SMSCs 的分化。在 SMSCs 中敲低 CDR1as 后,miR-27a-3p 差异上调。过表达 miR-27a-3p 降低了 SMSCs 的分化。通过 RNAhybrid 和荧光素酶实验,鉴定出 miR-27a-3p 可以调节 ANGPT1。我们发现,miR-27a-3p 与 CDR1as 呈负相关,并且在 SMSCs 分化过程中降低 ANGPT1 的表达水平。总之,我们的研究表明,siCDR1as 通过 miR-27a-3p 下调 ANGPT1 mRNA 抑制成肌细胞分化。