LincRNA Competitive Binding MiRNA To Regulate The Expression Of MRNA In Mouse Neural Development

Large intergenic non-coding RNAs are non-coding RNA with lengths more than 200 nt,it involved in many biological processes,now lncRNA has become one of the hot research in biological field.The available research results indicate that lincRNA can play an important role in brain development.In this thesis bioinformatics methods were employed to investigate the mouse neural stem cell transcriptome during the differentiation,and try to reveal lincRNA's function during differentiation.The main content of this paper are as follows:Firstly,We obtained RNA-seq(poly+A)from Wild type and Smek knockdown mouse neural stem cell(NSC)in the time course of 0 day,2 days and 6 days of differentiation with three.We identified the differential expression mRNAs and lincRNAs during differentiation by Tophat,Cufflinks,Cuffmerge,and Cuffdiff.We found 5271 mRNAs and 173 lincRNAs were differentially expressed between the 0 day and 2 days of NSC differentiation,and 1581 mRNAs and 71 lincRNAs were differentially expressed between 2 days and 6 days of NSC differentiation.We predicted the miRNA's target sites in lincRNAs,and found that this lincRNA target sites count was positive correlated with the GC content and negative correlative with the minimum free energy(MFE)of lincRNA using statistical analysis.Secondly,we obtained the data of miRNA's mRNA target gene validated by experiment in the database of miRTarBase,and the interaction between lincRNA and mi RNA predicted by miRanda,thus we obtained lincRNAs which can regulate the mRNA expression mediated by miRNA,and these lincRNAs and mRNA were found to co-expressed during NSC differentiation.We found that mRNA Bicd1,Irgq can promote the NSC's differentiation,and their sponge lincRNA Gm26888-201,6330403K07Rik-001 have the similar function to promote NSC's differentiation.By constructing lincRNA-miRNA-mRNA the gene regulatory network,we found that these lincRNAs Gm11549-001,643056201Rik-202,Trp53cor1-001 could be the factor of maintenance of neural stem cell,and lincRNAs Miat-002,Gm26888-201 could promote NSC's differentiation.GO enrichment analysis also revealed that lincRNA can play a role mainly in the neural differentiation,cell metabolism,and Wnt signaling pathway during NSC's differentiation process.Finally,we analyzed the gene Smek's function in NSC's differentiation process.By employing GO enrichment analysis tools DAVID and BINGO,we found that Smek can act in the limbic system development and transcription factor activity.And there was a close connection between lincRNA and gene Smek.When knocking down gene Smek,the lincRNA's(Gm26578-201,Gm27032-001,Mir124a-1hg-001)were regulated by expression down in the 2 days of NSC differentiation,the down-regulated lincRNA's sponge mRNAs can promote the differentiation of NSC,revealed that knockdown gene Smek can inhibit some lincRNA expression,and maintain neural stem cell.