牛病毒性腹泻病毒复制相关宿主细胞环状RNA的筛选

Screening of Host Cell CircRNA Related to Replication of Bovine Viral Diarrhea Virus

为了明确环状RNA(circRNA)在牛病毒性腹泻病毒(BVDV)复制中的作用,用BVDV NADL感染牛肾细胞(MDBK)并进行高通量测序分析,结果获得大量差异表达circRNA,对高通量测序数据进行筛选,共获得了12个上调的circRNA,通过RNase R消化、荧光定量PCR验证及通路分析,初步认为circ-0027407具有抑制病毒复制的可能性。在RNA干扰试验中,设计的干扰片段显著降低了circ-0027407的表达量,降低幅度约为66.1%,使用干扰载体转染MDBK细胞后感染病毒,发现干扰circ-0027407会显著增加病毒复制,提高了约113.2%。说明circ-0027407对BVDV感染有抑制作用。

The aim of research was to study the circular RNA(circRNA)in bovine viral diarrhea virus(BVDV).Mafia-Darby bovine kidney cells(MDBK)were infected with BVDV NADL,and Qualcomm was used.The quantitative sequencing analysis showed that a large number of differentially expressed circRNAs were obtained.High-throughput sequencing data were screened.A total of 12 potential circRNAs were obtained,which were verified by RNase R digestion and real-time PCR,and the pathway analysis was carried out,which initially considered that circ-0027407 has the potential to inhibit viral replication.In the RNA interference experiment,the designed interference fragment significantly reduced the expression of circ-0027407 by about 66.1%.After transfecting the MDBK cells with the interference vector,the virus was found to interfere with circ-0027407,which significantly increased virus replication by about 113.2%.These results demonstrated the inhibitory effect of circ-0027407 on BVDV infection.