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鉴定和共表达分析与奥汗细毛羊肌内脂肪沉积相关的长非编码 RNA 和 mRNAs。

Identification and co-expression analysis of long noncoding RNAs and mRNAs involved in the deposition of intramuscular fat in Aohan fine-wool sheep.

机构信息

College of Animal Science and Technology, Qingdao Agricultural University, Qingdao, 266109, China.

Qufu Animal Husbandry and Veterinary Technical Service Center, Qufu, 273100, China.

出版信息

BMC Genomics. 2021 Feb 1;22(1):98. doi: 10.1186/s12864-021-07385-9.

Abstract

BACKGROUND

Intramuscular fat (IMF) content has become one of the most important indicators for measuring meat quality, and levels of IMF are affected by various genes. Long non-coding RNAs (lncRNAs) are widely expressed non-coding RNAs that play an important regulatory role in a variety of biological processes; however, research on the lncRNAs involved in sheep IMF deposition is still in its infancy. Aohan fine-wool sheep (AFWS), one of China's most important meat-hair, dual-purpose sheep breed, provides a great model for studying the role of lncRNAs in the regulation of IMF deposition. We identified lncRNAs by RNA sequencing in Longissimus thoracis et lumborum (LTL) samples of sheep at two ages: 2 months (Mth-2) and 12 months (Mth-12).

RESULTS

We identified a total of 26,247 genes and 6935 novel lncRNAs in LTL samples of sheep. Among these, 199 mRNAs and 61 lncRNAs were differentially expressed. We then compared the structural characteristics of lncRNAs and mRNAs. We obtained target genes of differentially expressed lncRNAs (DELs) and performed enrichment analyses using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). We found that target mRNAs were enriched in metabolic processes and developmental pathways. One pathway was significantly enriched, namely tight junction. Based on the analysis of critical target genes, we obtained seven candidate lncRNAs that potentially regulated lipid deposition and constructed a lncRNA-mRNA co-expression network that included MSTRG.4051.3-FZD4, MSTRG.16157.3-ULK1, MSTRG.21053.3-PAQR3, MSTRG.19941.2-TPI1, MSTRG.12864.1-FHL1, MSTRG.2469.2-EXOC6 and MSTRG.21381.1-NCOA1. We speculated that these candidate lncRNAs might play a role by regulating the expression of target genes. We randomly selected five mRNAs and five lncRNAs to verify the accuracy of the sequencing data by qRT-PCR.

CONCLUSIONS

Our study identified the differentially expressed mRNAs and lncRNAs during intramuscular lipid deposition in Aohan fine-wool sheep. The work may widen the knowledge about the annotation of the sheep genome and provide a working basis for investigating intramuscular fat deposition in sheep.

摘要

背景

肌内脂肪(IMF)含量已成为衡量肉质的最重要指标之一,而 IMF 水平受多种基因的影响。长链非编码 RNA(lncRNA)是一种广泛表达的非编码 RNA,在多种生物过程中发挥着重要的调控作用;然而,关于 lncRNA 参与绵羊 IMF 沉积的研究仍处于起步阶段。奥汗细毛羊(AFWS)是中国最重要的肉毛两用绵羊品种之一,为研究 lncRNA 在调节 IMF 沉积中的作用提供了很好的模型。我们在绵羊 2 月龄(Mth-2)和 12 月龄(Mth-12)的背最长肌(LTL)样本中通过 RNA 测序鉴定了 lncRNA。

结果

我们共鉴定了绵羊 LTL 样本中的 26247 个基因和 6935 个新的 lncRNA。其中,199 个 mRNA 和 61 个 lncRNA 差异表达。然后,我们比较了 lncRNA 和 mRNA 的结构特征。我们获得了差异表达 lncRNA(DEL)的靶基因,并使用基因本体论(GO)和京都基因与基因组百科全书(KEGG)进行了富集分析。我们发现靶 mRNA 富集在代谢过程和发育途径中。有一个途径显著富集,即紧密连接。基于对关键靶基因的分析,我们获得了七个潜在调节脂质沉积的候选 lncRNA,并构建了一个包含 MSTRG.4051.3-FZD4、MSTRG.16157.3-ULK1、MSTRG.21053.3-PAQR3、MSTRG.19941.2-TPI1、MSTRG.12864.1-FHL1、MSTRG.2469.2-EXOC6 和 MSTRG.21381.1-NCOA1 的 lncRNA-mRNA 共表达网络。我们推测这些候选 lncRNA 可能通过调节靶基因的表达发挥作用。我们随机选择了 5 个 mRNA 和 5 个 lncRNA 通过 qRT-PCR 验证测序数据的准确性。

结论

本研究鉴定了奥汗细毛羊肌肉内脂质沉积过程中差异表达的 mRNA 和 lncRNA。这项工作可能拓宽了对绵羊基因组注释的认识,并为研究绵羊肌肉内脂肪沉积提供了工作基础。

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