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猪骨骼肌发育过程中长非编码 RNA 和 mRNAs 的分析与功能研究

Profiling and Functional Analysis of Long Noncoding RNAs and mRNAs during Porcine Skeletal Muscle Development.

机构信息

Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China.

Institute of Animal Husbandry and Veterinary, Guizhou Academy of Agricultural Science, Guiyang 550005, China.

出版信息

Int J Mol Sci. 2021 Jan 6;22(2):503. doi: 10.3390/ijms22020503.

Abstract

Gene transcripts or mRNAs and long noncoding RNAs (lncRNAs) are differentially expressed during porcine skeletal muscle development. However, only a few studies have been conducted on skeletal muscle transcriptome in pigs based on timepoints according to the growth curve for porcine. Here, we investigated gene expression in Qingyu pigs at three different growth stages: the inflection point with the maximum growth rate (MGI), the inflection point of the gradually increasing stage to the rapidly increasing stage (GRI), and the inflection point of the rapidly increasing stage to the slowly increasing stage (RSI). Subsequently, we explored gene expression profiles during muscle development at the MGI, GRI and RSI stages by Ribo-Zero RNA sequencing. Qingyu pigs reached the MGI, GRI and RSI stages at 156.40, 23.82 and 288.97 days of age with 51.73, 3.14 and 107.03 kg body weight, respectively. A total of 14,530 mRNAs and 11,970 lncRNAs were identified at the three stages, and 645, 323 differentially expressed genes (DEGs) and 696, 760 differentially expressed lncRNAs (DELs) were identified in the GRI vs. MGI, and RSI vs. MGI, comparisons. Functional enrichment analysis revealed that genes involved in immune system development and energy metabolism (mainly relate to amino acid, carbohydrate and lipid) were enriched at the GRI and MGI stages, respectively, whereas genes involved in lipid metabolism were enriched at the RSI stage. We further characterized , an abundant lncRNA. The full-length sequence (316 nt) of lncRNA was determined by rapid amplification of cDNA ends (RACE). Subcellular distribution analysis by quantitative real-time PCR (qRT-PCR) revealed that is a cytoplasmic lncRNA. Binding site prediction and dual luciferase assay showed that lncRNA directly binds to microRNA 133a (miR-133a). Our findings provide the basis for further investigation of the regulatory mechanisms and molecular genetics of muscle development in pigs.

摘要

基因转录本或 mRNAs 和长非编码 RNA(lncRNAs)在猪骨骼肌发育过程中表现出差异表达。然而,基于猪生长曲线的时间点,仅有少数研究针对猪骨骼肌转录组进行了研究。在这里,我们在三个不同的生长阶段研究了青峪猪的基因表达:增长率最大的拐点(MGI)、逐渐增加阶段到快速增加阶段的拐点(GRI)和快速增加阶段到缓慢增加阶段的拐点(RSI)。随后,我们通过核糖体零 RNA 测序技术,在 MGI、GRI 和 RSI 阶段探索了肌肉发育过程中的基因表达谱。青峪猪在 156.40、23.82 和 288.97 日龄时达到 MGI、GRI 和 RSI 阶段,体重分别为 51.73、3.14 和 107.03 公斤。在这三个阶段共鉴定出 14530 个 mRNAs 和 11970 个 lncRNAs,在 GRI 与 MGI、RSI 与 MGI 的比较中,分别鉴定出 645 个、323 个差异表达基因(DEGs)和 696 个、760 个差异表达 lncRNAs(DELs)。功能富集分析表明,在 GRI 和 MGI 阶段,分别富集了参与免疫系统发育和能量代谢的基因(主要与氨基酸、碳水化合物和脂质有关),而在 RSI 阶段,富集了参与脂质代谢的基因。我们进一步对丰富的 lncRNA 进行了表征。通过快速扩增 cDNA 末端(RACE)确定了 lncRNA 的全长序列(316nt)。通过定量实时 PCR(qRT-PCR)的亚细胞分布分析显示,是一种细胞质 lncRNA。结合位点预测和双荧光素酶报告基因检测实验表明,lncRNA 直接与 microRNA 133a(miR-133a)结合。我们的研究结果为进一步研究猪肌肉发育的调控机制和分子遗传学提供了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/736c/7825455/da1143a33728/ijms-22-00503-g001.jpg

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