ArticleScientific reports2015
Development of a cell system for siRNA screening of pathogen responses in human and mouse macrophages.
Article in Scientific reports, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 23 citations in OpenAlex.
- Antisense oligonucleotide-based switches for conditional translation of synthetic mRNA.Molecular therapy. Nucleic acids · 2025Article
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- Evidence of collective influence in innate sensing using fluidic force microscopy.Frontiers in immunology · 2024Article
- Single-cell analysis reveals a weak macrophage subpopulation response to Mycobacterium tuberculosis infection.Cell reports · 2023Article
- Nicotinamide Antagonizes Lipopolysaccharide-Induced Hypoxic Cell Signals in Human Macrophages.Journal of immunology (Baltimore, Md. : 1950) · 2023Article
- Small-molecule screening identifies Syk kinase inhibition and rutaecarpine as modulators of macrophage training and SARS-CoV-2 infection.Cell reports · 2022Article
- A genome-wide screen uncovers multiple roles for mitochondrial nucleoside diphosphate kinase D in inflammasome activation.Science signaling · 2021Article
- Type I IFNs facilitate innate immune control of the opportunistic bacteria Burkholderia cenocepacia in the macrophage cytosol.PLoS pathogens · 2021Article
- XL-DNase-seq: improved footprinting of dynamic transcription factors.Epigenetics & chromatin · 2019Article
- IFIT1 Exerts Opposing Regulatory Effects on the Inflammatory and Interferon Gene Programs in LPS-Activated Human Macrophages.Cell reports · 2018Article
- A poly(beta-amino ester) activates macrophages independent of NF-κB signaling.Acta biomaterialia · 2018Article
- Measurement of NF-κB Activation in TLR-Activated Macrophages.Methods in molecular biology (Clifton, N.J.) · 2018Article
- Systematic Investigation of Multi-TLR Sensing Identifies Regulators of Sustained Gene Activation in Macrophages.Cell systems · 2017Article
- Knockdown of Host Antioxidant Defense Genes Enhances the Effect of Glucantime on Intracellular Leishmania braziliensis in Human Macrophages.Antimicrobial agents and chemotherapy · 2017Article
- Genome-wide siRNA screen of genes regulating the LPS-induced NF-κB and TNF-α responses in mouse macrophages.Scientific data · 2017Article
- Genome-wide siRNA screen of genes regulating the LPS-induced TNF-α response in human macrophages.Scientific data · 2017Article
- Overcoming Gene-Delivery Hurdles: Physiological Considerations for Nonviral Vectors.Trends in biotechnology · 2016Review
- Comprehensive RNAi-based screening of human and mouse TLR pathways identifies species-specific preferences in signaling protein use.Science signaling · 2016Article
- Assay Development for Image-Based Quantification of Intracellular Bacterial Replication and Analysis of the Innate Immune Response to Infection.Assay and drug development technologies · 2015Article
- Article
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Authors and funding
9 authors at 3 institutions in 1 country.
Funding
Abstract
Macrophages play a critical role in the innate immune response to pathogen infection, but few tools exist for systematic dissection of these responses using modern genome-wide perturbation methods. To develop an assay platform for high-throughput analysis of macrophage activation by pathogenic stimuli, we generated reporter systems in human and mouse macrophages with dynamic readouts for NF-κB and/or TNF-α responses. These reporter cells show responsiveness to a broad range of TLR ligands and to gram-negative bacterial infection. There are significant challenges to the use of RNAi in innate immune cells, including efficient small RNA delivery and non-specific immune responses to dsRNA. To permit the interrogation of the macrophage pathogen response pathways with RNAi, we employed the stably expressed reporter genes to develop efficient siRNA delivery protocols for maximal target gene silencing with minimal activation of the innate macrophage response to nucleic acids. We demonstrate the utility of these macrophage cell systems for siRNA screening of pathogen responses by targeting components of the human and mouse TLR pathways, and observe species-specific perturbation of signaling and cytokine responses. Our approach to reporter cell development and siRNA delivery optimization provides an experimental paradigm with significant potential for developing genetic screening platforms in mammalian cells.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.