Fkbp51基因敲除对小鼠肝脏转录组基因可变剪接的影响

摘要

目的 通过分析Fkbp51基因敲除(knock out,KO)与野生型(wild type,WT)小鼠肝脏表达谱,研究Fkbp51基因敲除对肝脏组织基因可变剪接的影响.方法 利用二代测序对Fkbp51 KO与WT小鼠肝脏进行表达谱测序,用TopHat对RNA测序结果进行可变剪接分析,筛选出KO与WT小鼠肝组织中差异的内含子保留(intron retetion,RI)和外显子跳跃(exon skipping,SE).通过在线工具DAVID对这些差异可变剪接体进行基因功能 (gene ontology, GO)和代谢通路(kyoto encyclopedia of genes and genomes,KEGG)富集分析,同时用NCBI 基因数据库对这些基因进行注释.结果 (1)Fkbp51缺失可导致小鼠肝脏mRNA可变剪接发生变化;(2)Fkbp51基因敲除造成小鼠肝脏mRNA可变剪接表达量的变化;(3)通过GO与KEGG分析,我们发现这些发生差异可变剪切的基因主要与脂肪相关衍生物的代谢、免疫、胆汁酸分泌等通路相关.(4)与差异内含子保留相关的基因主要与肌动蛋白细胞骨架调控,氨基酸及其衍生物代谢相关.结论 Fkbp51基因敲除能够改变基因组中mRNA的可变剪切,进而影响小鼠肝脏的代谢功能.%Objective The purpose of this study is to understand the influence of Fkbp51 gene knockout on alternative splicing of mRNA in liver tissues.Methods mRNAs of liver from both Fkbp51 knockout(KO) and wild type(WT) mice were isolated.mRNA expression profiling was performed using RNA-seq reads.The mRNA reads produced from RNA-seq was analysed by TopHat for alternative splicing.Exon skipping and intron retetion were identified according to alignment analyses.Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were analyzed using online tools DAVID.All genes which had differently expressed alternative splicing between WT and KO were annotated using NCBI database.Results (1) Fkbp51 KO can introduce new mRNA alternative splicing;(2) Fkbp51 KO also cause mRNA alternative splicing expression change;(3) According to GO and KEGG analysis, it was found that the mRNA alternative splicing changes mainly enriched in fat metabolism, immune, bile acid secretion, and PPAR signaling pathway etc.(4) The genes which have different intron retetion events are mainly involved in regulation of actin cytoskeleton organization and amino acids metabolism.Conclusions Fkbp51 gene knockout can cause the change of mRNA alternative splicing, and thereby affect the metabolism of liver in mice.