Specifically, we focused on the single miRNA accounting for almost 50% of miRNAs constitutively expressed in human T cells, miR-150-5p (hereafter miR-150).

In this study, we took advantage of an integrated approach combining quantitative miRNA analysis, unbiased genomics, and genome editing to identify miRNAs highly expressed in primary human T lymphocytes, analyze the regulatory logic underpinning their expression, and finally characterize mRNA target regulation and their functional impact. Examples in this group include miR-155 and miR-146a, which are responsible for enhancing and attenuating T-cell responses, respectively. Finally, modestly expressed but inducible miRNAs may dynamically reach intracellular concentrations relevant in the modulation of T-cell activation. By contrast, miRNAs that are expressed at very low levels are highly unlikely to reach the concentrations required to exert biological functions. Abundant miRNAs that are down-regulated after stimulation may be involved in restraining T-cell activation, as shown in the case of miR-125b, which is required to maintain the naive state of human T cells. The quantitative analysis of miRNA expression in different T-cell subsets and in response to T cell receptor (TCR) triggering may thus provide clues on the functional impact of individual miRNAs on T-cell responses. The effect of any given miRNA is dependent on its expression level relative to that of its targets and also on the specific context and cell-specific usage of target sites in the 3РІР‚Р† untranslated region (UTR) of mRNAs, resembling the cell typeРІР‚вЂњspecific regulation of gene expression mediated by transcription factors. Through their ability to target a variety of mRNAs and regulate their translation and stability, microRNAs (miRNAs) modulate all aspects of the biology of T lymphocytes, including cell differentiation, activation, and proliferation. Reverse transcription quantitative PCR sgRNA, The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Р”В¬ompeting interests: The authors have declared that no competing interests exist.Р”Р„ssay for transposase accessible chromatin and sequencing BrdU,Р”В¬arboxyfluorescein succinimidyl ester ChIP-seq,Р”В¬hromatin immunoprecipitation sequencing crRNP,Р”Вatabase of Disordered Protein Predictions DICE, All other relevant data are within the paper and its Supporting Information files S1 Data (Data Table) and S4 Fig (Raw images).Р”Р‡unding: This work was supported by the Swiss National Science Foundation grant 31003A_175569 ( the NCCR РІР‚СљRNA & Disease', the Novartis Foundation for medical-biological Research and the Ceresio Foundation (all to SM). This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Р”Вata Availability: Datasets are deposited at GEO with accession number GSE161100. Received: ApAccepted: JanuPublished: February 10, 2022Р”В¬opyright: Р’В© 2022 Chirichella et al. Oliver, ChildrenРІР‚в„ўs Hospital of Philadelphia and The University of Pennsylvania School of Medicine, UNITED STATES PLoS Biol 20(2):Р”Р„cademic Editor: Paula M. (2022) RFX transcription factors control a miR-150/PDAP1 axis that restrains the proliferation of human T cells. Overall, using an integrated approach involving quantitative analysis, unbiased genomics, and genome editing, we identified RFX factors, miR-150, and the PDAP1 RBP as the components of a regulatory axis that restrains proliferation of primary human T lymphocytes.Р”В¬itation: Chirichella M, Bianchi N, DР•С•afo E, Foli E, Gualdrini F, Kenyon A, et al. PDAP1 acted as an RNA-binding protein (RBP), and its CRISPR/Cas-9РІР‚вЂњmediated deletion revealed that it prominently contributed to the regulation of T-cell proliferation. By performing miRNA pull-down and sequencing experiments, we identified PDGFA-associated protein 1 (PDAP1) as one main target of miR-150 in human T lymphocytes. Constitutive MIR150 gene expression required the RFX family of transcription factors, and its activation-induced down-regulation was linked to their reduced expression. We found that in primary human memory T helper lymphocytes, miR-150 was the most abundantly expressed miRNA, and its expression decreased drastically upon activation, suggesting regulatory roles. However, what are the mRNA targets regulated by miRNAs and how miRNAs are transcriptionally regulated themselves remain for the most part unknown. Within the immune system, microRNAs (miRNAs) exert key regulatory functions.

Cb rfx 150