自然反义转录物Lmo4as对神经系统重要编码基因Lmo4的调节作用

The natural antisense transcript,Lmo4as,negatively regulates the same locus cortical coding gene Lmo4

目的探索小鼠神经系统重要编码基因Lmo4基因座位上自然反义转录产物Lmo4as对该编码基因的表达调控功能。方法 1)使用c DNA末端快速扩增(RACE)与RT-PCR技术,验证Lmo4as是否具有多聚腺苷酸(poly A)尾,克隆Lmo4as的全长序列;2)使用实时定量PCR和原位杂交技术确定Lmo4as及相应编码基因的时空表达谱;3)使用荧光素酶报告基因实验在体外验证Lmo4as对Lmo4是否具有调控作用。结果 1)Lmo4as是一个具有多聚腺苷酸(poly A)尾的非编码RNA转录本,获得其RNA全长信息;2)Lmo4as与相应编码基因转录Lmo4的丰度在时间上具有协同性,而在空间表达上具有明显不同的分布;3)Lmo4as对Lmo4在转录后水平具有负性调节作用,另外miR-124也能够抑制Lmo4的表达。结论编码基因Lmo4基因座位上自然反义转录产物Lmo4as对其转录后水平的负性调节与microRNA的调节共同发挥作用。

Objective Investigation of the cis regulatory functional role of natural antisense transcript （NAT） in expression of same genome locus coding gene, Lmo4. Methods Rapid amplification of cDNA ends （ RACE ）, RT- PCR, real time qRT-PCR, in situ hybridization and luciferase reporter assay were employed to capture the full length of the NAT, to display the spatio-temporal patterns of coding gene and noncoding RNA in the development of embryonic brain, and to analyze the regulatory role of NAT in the expression of Lmo4, respectively. Results The results showed that Lmo4as was a noncoding RNA transcript with a poly-A tail, and the full length sequence of this NAT transcript was cloned. Moreover, it was shown that Lmo4as function as a negative regulator on the expression of Lmo4 at the post-transcriptional level, similar with a microRNA functional role, miR-124. Conclusions The NAT Lmo4as functions as a cis repressor on the coding gene Lmo4 at post-transcriptional level.