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Effect of miR-144/miR451 expression on TC-1 lung epithelial cell responses to influenza infection for 24 hours [RT-PCR]

ABSTRACT: Antiviral responses must be regulated to rapidly defend against infection while minimizing inflammatory damage, but the mechanisms for establishing the magnitude of response within an infected cell are not well understood. miRNAs are small non-coding RNAs that negatively regulate protein levels by binding target sequences on their cognate mRNA. We profiled microRNA expression in the lungs of mice infected for 24 h with Influenza A/PR/8/34 to identify microRNAs that may regulate host response to influenza infection. Mice were infected intranasally with 10e5 pfu Influenza A/PR/8/34. Lungs were isolated after 24 hours and total RNA extracted using Trizol. Equivalent quantities of RNA from 3 infected mice were pooled.

ORGANISM(S): Mus musculus

SUBMITTER: madhumati gundapuneni

PROVIDER: E-GEOD-31956 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Effect of miR-144/miR451 expression on TC-1 lung epithelial cell responses to influenza infection for 24 hours

Project description:This SuperSeries is composed of the following subset Series: GSE31955: Effect of miR-144/miR451 expression on TC-1 lung epithelial cell responses to influenza infection for 24 hours [Expression] GSE31956: Effect of miR-144/miR451 expression on TC-1 lung epithelial cell responses to influenza infection for 24 hours [RT-PCR] Refer to individual Series

2012-07-31 | E-GEOD-31957 | biostudies-arrayexpress

Effect of miR-144/miR451 expression on TC-1 lung epithelial cell responses to influenza infection for 24 hours [RT-PCR]

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Effect of miR-144/miR451 expression on TC-1 lung epithelial cell responses to influenza infection for 24 hours [Expression]

Project description:Antiviral responses must be regulated to rapidly defend against infection while minimizing inflammatory damage, but the mechanisms for establishing the magnitude of response within an infected cell are not well understood. miRNAs are small non-coding RNAs that negatively regulate protein levels by binding target sequences on their cognate mRNA. Here we identify miR-144 as a negative regulator of the host antiviral response. Ectopic expression of miR-144 resulted in increased replication of three RNA viruses, influenza, EMCV, and VSV, in primary mouse lung epithelial cells. To elucidate the mechanism whereby miR-144 increases influenza replication within lung epithelial cells, TC-1 cells stably over-expressing miR-144 were infected with influenza A for 24 hours and the transcriptional profile was compared with those of infected control cells. This systems biology approach identified the transcriptional network regulated by miR-144 and demonstrate that it controls the TRAF6/IRF7 antiviral response by post-transcriptionally suppressing TRAF6 levels. In vivo ablation of miR-144 reduced influenza replication within the lung. TC-1 lung epithelial cells stably expressing miR144+miR451 or control vector were unstimulated (n=1) or infected with Influenza A/PR/8/34 (MOI=5) for 24 hours (n=3).

2012-07-31 | E-GEOD-31955 | biostudies-arrayexpress

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Project description:C57BL/6J (WT) mice were infected with influenza A virus PR/8/34 or mock infected with phosphate buffered saline (PBS). 18 hours later lung tissue was harvested for transcriptional profiling.

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microRNA signatures of iPSCs and endoderm-derived tissues

Project description:MicroRNAs (miRNAs), small non-coding RNAs that fine-tune gene expression, play multiple roles in the cell, including cell fate specification. We have analyzed the differential expression of miRNAs during fibroblast reprogramming into induced pluripotent stem cells (iPSCs) and endoderm induction in iPSCs upon treatment with high concentrations of Activin-A in reduced serum. During reprogramming, adult mouse fibroblasts are converted into cells that resemble embryonic stem cells (ESCs) according to standard molecular and functional assays for pluripotency. The reprogrammed iPSCs assume an ESC-like miRNA signature, marked by the strong induction of pluripotency clusters miR-290-295 and miR-302/367 and conversely the downregulation of the let-7 family. On the other hand, endoderm induction in iPSCs results in the upregulation of 13 miRNAs. Given that the liver and the pancreas are common derivatives of the endoderm, the comparison of the expression levels of these 13 upregulated miRNAs with those in hepatocytes and pancreatic islets suggests a trend of miRNA upregulation in the endoderm tending towards an islet phenotype rather than that of a hepatocyte. These observations provide insights into how differentiation may be guided more efficiently towards the endoderm and further into the liver or pancreas. Moreover, we also report novel miRNAs enriched for each of the cell types analyzed. Stemloop RT-qPCR gene expression profiling. REPROGRAMMING: Differentially expressed miRNAs were determined between iPSCs (n=5 clones) and parent tail-tip fibroblasts (n=5) using mESCs R1 (n=3) and D3 (n=3). DIFFERENTIATION: Differentially expressed miRNAs were also analyzed in two iPSC clones upon treatment with Activin-A (n=2 each), and between primary mouse hepatocytes (n=3) and pancreatic islets (n=3).

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