Expression Profiles Changes Of LncRNA And MRNA And Their Function Prediction In Human Cells Infected With Fixed Rabies Virus PM Strain

Rabies is a highly lethal zoonotic infectious disease caused by rabies virus that can be effectively prevented by vaccination before and after exposure,but almost 100% of deaths occur once the disease take place.Although the researcher’s unremitting efforts have made great progress in the molecular biological characteristics and pathogenic mechanism of rabies virus,the problem of incurable rabies disease remains unresolved.In recent years,with the development of high-throughput sequencing technology and genomics,long non-coding RNAs(lncRNAs)have become a research hotspot.More and more studies have shown that LncRNA plays an important role in viral infection.The purpose of this study was to investigate the changes of lncRNA and mRNA expression profiles and to predict the function of human rabies virus-infected PM strains after infection by RNA sequencing,and to provide a direction for the study of the pathogenesis of rabies virus.In this study,human A549 cells were selected as hosts for PM strain infection.The total RNA was extracted after infection with multiple MOI=1,48 hours after infection,and sent to the service company for RNA quality detection,database construction,high-throughput sequencing and subsequent analysis,and the experimental group was established.Three independent experiments were performed in each of the control and the control group,and a total of 6 samples were used to verify the transcript.After the high-throughput sequencing,the original off-machine data was filtered and analyzed.The results showed that the sequencing quality was high and met the requirements.Transcripts were spliced for high-quality filtered data,and transcripts were classified into mRNA,known lncRNA,and other unknown transcripts based on the alignment of the transcript with the reference genome.Direct expression analysis of mRNA and known lncRNA and functional analysis of differential genes;other unknown transcripts were first screened according to some basic conditions of lncRNA,and then analyzed by 4 coding potential analysis software,the last 6 samples A total of 16284 novel lncRNA were obtained.The results of cluster analysis of sample expression showed that the mRNA and LncRNA expression profiles of the PM strain infected group and the PBS control group changed significantly.Functional analysis of GO and KEGG differentially expressing mRNA and lncRNA revealed that there were more than 1000 biological processes for differentially expressed mRNA enrichment,and 80 pathways for enrichment;more than 2,000 biological processes for differential expression of lncRNA enrichment.There were 170 pathways,indicating that the changes in lncRNA expression profiles induced by PM strain infection were much higher than those of mRNA expression profiles.However,the biological processes that differentially express lncRNA and mRNA enrichment were consistent,including cell processes,developmental processes,cell differentiation,signal regulation processes,energy metabolism,glucose metabolism,disease processes,and immune defense.Functional analysis showed that although the host cell function changes caused by PM strain infection were very extensive,these changes were not uniform,and were more focused on the biological functions related to antiviral responses such as cellular immunity,inflammatory response,autophagy and apoptosis.Finally,we selected 4 significantly up-regulated and 4 significantly down-regulated qRT-PCR validations from differentially expressed lncRNA.The results were consistent with the sequencing results,which confirmed the accuracy of high-throughput sequencing.This study showed that the infection of rabies virus PM strain significantly changed the expression profile of lncRNA and mRNA in A549 cells,indicating that the infection of PM strain caused a wide change in the biological function of human cells.Although functional analysis has shown that the biological processes of host cells have changed in all aspects,the changes in biological functions related to antiviral responses such as cellular immunity,inflammatory response,autophagy and apoptosis were particularly prominent.In this study,the expression profile of lncRNA in host cells infected by PM strain reveals the regulation mechanism of lncRNA regulation of rabies virus infection and rabies virus escape host immunity during rabies virus-infected cells,which lays a foundation for further prevention and treatment of rabies basis.