The Study On Expression Of LncRNA-H13 In Embryo Development And A Preliminary Study Of Its Function

Lnc RNA refer to RNA molecules greater than 200 bp in length,with no protein coding ability, and have similar structure with the encoding gene, and exist a phenomenon of post-transcriptional splicing. The spatio-temporal specificity in Lnc RNA show that such RNA are associated with a variety of activities related to life, and the function of Lnc RNA are often known by regulating the expression of other genes, so far the studies about the function of Lnc RNA have mainly been focused on knockout and overexpression.This paper is based on the long non-coding RNA associated with mouse embryonic growth and development, which are predicted by RNA-seq data, and a preliminary analysis to verify its existence, and then selected Lnc-H13 as the final study as it have a relatively higher expression. Lnc-H13 is located on mouse chromosome 2, containing only one exon region. Bioinformatics forecast the full-length of Lnc-H13 is 7122 bp. Hic data showed that Lnc-H13 acts on Mcts2 and H13. Therefore, We analyzed the Lnc-H13 expression patterns in mouse embryos and make a preliminary explore about its function.Firstly, We analyzed Lnc-H13 in mouse embryonic quantification and localization by using the technologies of q RT-PCR and in situ hybridization. The results showed that in whole embryos Lnc-H13 expression level gradually increased with the development of embryos and reached a peak of expression in E15.5 day. E10.5 embryos in situ hybridization showed that Lnc-H13 is related to the formation of nervous system in early embryonic development. E12.5 day embryos slice hybridization reveled that Lnc-H13 expressed in brain and lungs. With the embryonic development to med-term, Lnc-H13 is widely expressed in the brain, tongue, heart, lungs, kidneys and other vital organs. As Lnc-H13 has been always expressing in brain tissue, We analyzed the Lnc-H13 expression in different stages of development in the mouse brain, the results showed that with the development of embryos, the expression of Lnc-H13 in the brain increased firstly and then decreased, but before the born of mouse, Lnc-H13 expression had a slight fluctuation. Based on this phenomenon, We analyzed the localization of Lnc-H13 in postnatal mouse brain, the results showed that the gene is ubiquitously expressed in P0,P10 and P30 of the olfactory bulb and cerebellum. Above these, we can concluded that the Lnc-H13 gene is involved in the process of embryonic development, and plays an important role in the development of nervous system. Finally, We used the CRISPR/Cas9 technology made a gene knockout for the Lnc-H13 in NIH3T3 cells,It has been detected that when Lnc-H13 expression was down regulated,the expression level of target gene was not statistically significance.In summary, We analyzed the Lnc-H13 gene in mouse embryonic expression and postnatal expression in the brain and its functions in this study.