Effect Of Fkbp51 Gene Knockout On Variable Splicing Of Mouse Liver Transcriptome Gene

【Objective】Mstn(myostatin) is a negative regulator of skeletal muscle growth, which not only regulates the number and size of skeletal muscle, but also plays an important role in the regulation of lipogenesis and metabolism. Although there are Mstn gene knockout mice and large animals, large animals exist defects in the long period of reproduction, many physiological characteristics of mice are far apart from human, and it is necessary to construct Mstn gene knockout rats to make up for deficiencies of both mice and large mammals. Using Mstn KO rats as a mode to thoroughly study the mechanism of Mstn gene in the metabolism。[Methods] In this paper, using zinc finger nuclease(ZFN) technology targeted a site on Mstn exon 1, resulting in a nucleotide deletion and two nucleotides mutations. The frame shift mutation produced a premature termination codon, and finally we succeeded in constructing the Mstn gene knockout rats that only expressed the N-terminal 109 amino acids, not expressed the C-terminal fragment with biological activity.[Results] By phenotypic analysis of the rats, compared with littermate control rats, Mstn KO rats significantly put on weight and has a increased body size; The body muscle content increased, but grip force test revealed no fifference, suggesting that Mstn gene Knockout resulted in muscle content increased, but did not enhanced the strength of the muscle fibers; Mstn ko rat had reduced fat content through the analysis of fat content.【Conclusion】 These results indicate that Mstn gene knockout rats are successfully constructed, which will provide an important animal model for further study of the mechanism of Mstn gene in metabolism, treatment associated diseases and the development of animal husbandry.【 Objective]】The purpose of this study is to understand the influence of Fkbp51 knockout on alternative splicing of the orther genes by profiling gene expression of liver tissues of both Fkbp51KO and WT mice.[Methods] Liver’s mRNA of Fkbp51KO and WT mice were isolated and expression profiling was performed using RNA-seq.The reads produced from RNA-seq was analysed by TopHat for alternative splicing, and then filtered the differently expressed exon skipping and intron retetion, finally, Gene ontology and KEGG pathway enrichment was made by the use of online tool DAVID.On the same time,We make use of NCBI’s gene database to make annotation for those genes which was different in alternative splicing between KO and WT.【Results】 Fkbp51KO can cause new mRNA alternative splicing occurs; (2) Fkbp51 KO also cause mRNA alternative splicing expression changes;(3) Through GO and KEGG analysis, we found that these specific mRNA splicing changes mainly enriched in fat metabolismimmune, bile acid secretion, and PPAR signaling pathway and so forth.(4)We also found that those genes which has differently intron retetion mainly take part in regulation of actin cytoskeleton organization and metabolism of amino acids and their related derivatives.【Inclusions】Fkbp51 knockout can change the genome’s alternative splicing of mRNA, the occurrence of these mRNA splicing and expression changes is closely related to biological processes and pathways.