Mouse Cerebral Cortex Development Related To Screening And Identification Of Natural Antisense Gene Transcription

Noncoding RNA(NcRNA) has become one of hot point in today’s life science. Along with the important function of ncRNA be discovered we pay more attention to those small RNA molecules.Non-coding RNA formed a large network which plays a vital role in regulation of gene expression also has broad implications for biological growth and development, disease andother physiological processes.Long non-coding RNA (LncRNA) refers to a class of gene transcripts greater than200nt, but can not encode protein.Long non-coding RNAs have an important function on various stages of growth and development.During embryonic development long non-coding RNAs can be caused allele silence the maintenance of epigenetic traits and also associated with multi-cellular animals development and cell differentiation. Natural antisense transcripts is anew type of long non-coding RNA which transcripted from the antisense strand.Some of them can code protein.In1976, Barrell found that the presence of NAT in virus then successively been confirmed in humans, mice, rats, chickens, drosophila, melanogaster, rice, arabidopsis, yeast,nematode and other various plants and animals.NATs are highly prevalent in a wide range of species—for example,around15%of human protein-encoding genes have an associated NAT. In eukaryotes,NATs are linked to gene expression through genomic imprinting, RNA interference, transcriptional and post-translational regulation, alternative splicing, X-chromosome inactivation and RNA editing affected the development of organisms and the occurrence of the disease. However, we still know little about the role of NAT in the nervous system development. The purpose of our research was to find nats which have relations with cortex development in the mouse brain through large-scale screening and identification at the same time explore the preliminary functions of nats.We selected63genes which associated with the mouse cerebral cortex development as candidates. These genes belong to transcription factor which play an important role in cortex formation or marker molecules. They are the key transcriptional factors during nervous system development especially cerebral cortex development.63genes were screened by http://natsdb.cbi.pku.edu.cn, the prediction results showed that31genes with potential natural antisense transcripts.We detected nats from tissues of adult and E12.5mouse brain,MEF cell line,NIH3T3cell line then identified them through strand-specific RT-PCR method.Finally,8nats were proved to be positive.In order to in-depth exploration, we use the method of rapid amplification of cDNA ends to amplify3’and5’ end of8NATs cDNA. We discovered the details sequence information about3’end of Id2-NAT,Mef2c-NAT, Foxgl-NAT and5’end of LMO4-nat, Ldb2-NAT, Foxg1-NAT.NATs function is closely related with temporal and spatial position. We use the in situ hybridization method to observe the expression of Lmo4, Ldb2, Foxg1and its corresponding antisense transcripts in the mouse cerebral cortex in order to detect the interaction between the NAT and its gene, and understand its role in development of the nervous system We found that expression pattern of Foxgl-NAT appears a collaborative trend with Foxg1.Then detected the expression of Foxg1and its NAT in the mouse brain at different time point including E12.5,E14.5,E16.5,E18.5, we proved the existence of such collaborative trend indifferent developmental time points,provided a basis to explore the NAT functionality.