Differentially expressed long noncoding RNAs in RAW264.7 macrophages during Brucella infection and functional analysis on the bacterial intracellular replication
Abstract Long noncoding RNAs (lncRNAs) are a group of functional RNA molecules without protein-coding potential and play vital roles in majority of biological processes. To date, the expression profiles of lncRNAs and their influence on Brucella replication in RAW264.7 cells are poorly understood. I...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-12-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-25932-6 |
| _version_ | 1811188204329500672 |
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| author | Xiang Guan Hai Hu Minxing Tian Hongxu Zhuang Chan Ding Shengqing Yu |
| author_facet | Xiang Guan Hai Hu Minxing Tian Hongxu Zhuang Chan Ding Shengqing Yu |
| author_sort | Xiang Guan |
| collection | DOAJ |
| description | Abstract Long noncoding RNAs (lncRNAs) are a group of functional RNA molecules without protein-coding potential and play vital roles in majority of biological processes. To date, the expression profiles of lncRNAs and their influence on Brucella replication in RAW264.7 cells are poorly understood. In this study, we performed high-throughput transcriptome analysis to investigate the differentially expressed lncRNAs associated with Brucella abortus S2308 infection. Of these, 8, 6, 130 and 94 cellular lncRNAs were differentially expressed at 4, 8, 24 and 48 h post-infection, respectively. Moreover, 1918 protein-coding genes are predicted as potential cis target genes of differentially expressed lncRNAs by searching protein-coding genes located at upstream and downstream of lncRNA loci on the chromosome DNA of Mus musculus. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses indicated that majority of lncRNA target genes were associated with B. abortus infection. Fourteen lncRNAs from transcriptome data were selected for qRT-PCR verification, confirming 13 were differentially expressed. Animal experiments revealed three were differentially expressed in vivo by qRT-PCR analysis. Furthermore, knockdown of LNC_000428 by CRISPR/dCas9 inhibition or Locked Nucleic Acids transfection downregulated Tnfrsf8 expression at mRNA level and increased Brucella intracellular replication. Thus, we provide a novel evidence that lncRNAs induced by Brucella-infection function on Brucella intracellular replication. |
| first_indexed | 2024-04-11T14:16:29Z |
| format | Article |
| id | doaj.art-4602769ef8f44481ada50d0cfb3beaab |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-11T14:16:29Z |
| publishDate | 2022-12-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-4602769ef8f44481ada50d0cfb3beaab2022-12-22T04:19:24ZengNature PortfolioScientific Reports2045-23222022-12-0112111410.1038/s41598-022-25932-6Differentially expressed long noncoding RNAs in RAW264.7 macrophages during Brucella infection and functional analysis on the bacterial intracellular replicationXiang Guan0Hai Hu1Minxing Tian2Hongxu Zhuang3Chan Ding4Shengqing Yu5Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS)Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS)Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS)Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS)Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS)Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS)Abstract Long noncoding RNAs (lncRNAs) are a group of functional RNA molecules without protein-coding potential and play vital roles in majority of biological processes. To date, the expression profiles of lncRNAs and their influence on Brucella replication in RAW264.7 cells are poorly understood. In this study, we performed high-throughput transcriptome analysis to investigate the differentially expressed lncRNAs associated with Brucella abortus S2308 infection. Of these, 8, 6, 130 and 94 cellular lncRNAs were differentially expressed at 4, 8, 24 and 48 h post-infection, respectively. Moreover, 1918 protein-coding genes are predicted as potential cis target genes of differentially expressed lncRNAs by searching protein-coding genes located at upstream and downstream of lncRNA loci on the chromosome DNA of Mus musculus. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses indicated that majority of lncRNA target genes were associated with B. abortus infection. Fourteen lncRNAs from transcriptome data were selected for qRT-PCR verification, confirming 13 were differentially expressed. Animal experiments revealed three were differentially expressed in vivo by qRT-PCR analysis. Furthermore, knockdown of LNC_000428 by CRISPR/dCas9 inhibition or Locked Nucleic Acids transfection downregulated Tnfrsf8 expression at mRNA level and increased Brucella intracellular replication. Thus, we provide a novel evidence that lncRNAs induced by Brucella-infection function on Brucella intracellular replication.https://doi.org/10.1038/s41598-022-25932-6 |
| spellingShingle | Xiang Guan Hai Hu Minxing Tian Hongxu Zhuang Chan Ding Shengqing Yu Differentially expressed long noncoding RNAs in RAW264.7 macrophages during Brucella infection and functional analysis on the bacterial intracellular replication Scientific Reports |
| title | Differentially expressed long noncoding RNAs in RAW264.7 macrophages during Brucella infection and functional analysis on the bacterial intracellular replication |
| title_full | Differentially expressed long noncoding RNAs in RAW264.7 macrophages during Brucella infection and functional analysis on the bacterial intracellular replication |
| title_fullStr | Differentially expressed long noncoding RNAs in RAW264.7 macrophages during Brucella infection and functional analysis on the bacterial intracellular replication |
| title_full_unstemmed | Differentially expressed long noncoding RNAs in RAW264.7 macrophages during Brucella infection and functional analysis on the bacterial intracellular replication |
| title_short | Differentially expressed long noncoding RNAs in RAW264.7 macrophages during Brucella infection and functional analysis on the bacterial intracellular replication |
| title_sort | differentially expressed long noncoding rnas in raw264 7 macrophages during brucella infection and functional analysis on the bacterial intracellular replication |
| url | https://doi.org/10.1038/s41598-022-25932-6 |
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