Identification And Analysis Of Long Chain Non - Coding RNA Associated With Nbn ^CNS-del Mouse Cerebellar Defects

Nijmegen breakage syndrome (NBS) is an autosomal chromosomal breakage syndrome which is caused by hypomorphic mutation in NBS1 and characterized with microcephaly. Neuronal inactivation of Nbn in mice results in a combination of neurological anomalies characteristic of NBS, especially cerebellar defects. To identify novel long non-coding RNA (lncRNA) involved in cerebellar neurogenesis of neuronal specific Nbn-deficient (NbnCNS-del) mice, microarray analysis was performed to identify differentially expressed lncRNAs between wild type (NbnCNS-Ctr) and NbnCNS-del mice at p7 & p28. Two lncRNAs, Gml5577 and 1700020I14Rik, were chosen for further study including validation of expression profiles, subcellular fractio nation, and functional studies by using the knockout mouse model, primary cerebellar culture, P19 and Neuro-2a cell lines.Our results show that Gm15577 is specifically expressed in mice cerebellum in a developmentally regulated manner, which can be abolished upon Nbn-deficiency. It locates in the intronic region of Negrl in a reversed orientation. Gm15577 modulates the RNA expression of Negrl, Shh and β-catenin. Human NEGR1 has a distinct expression pattern between normal and medulloblastoma patients.1700020114Rik, mainly exists in cytoplasm and be highly expressed in cerebellum. During differentiation of primary cerebellar progenitor cells,1700020I14Rik exhibits the same expression pattern as neuronal marker β Ⅲ-tubulin. Moreover,1700020114Rik plays an important role in promoting RA-induced differentiation of P19 cells.1700020I14Rik shows a relatively higher conservation among different vetebrates, its human ortholog has been identified to be OIP5-AS1. As predicted by miRcode, OIP5-AS1 has many potential miRN As-binding sites, especially within a fragment highly conserved between 1700020I14Rik and OIP5-AS1 indicting a possibility of 1700020I14Rik as miRN A-sponge to execute its functions in promoting neuronal differentiation. Thus, our study reveals an important role for Gm15577 and 1700020114Rik in neurogenisis.