ArticleNature protocols2020
Rapid identification of human mast cell degranulation regulators using functional genomics coupled to high-resolution confocal microscopy.
Article in Nature protocols, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed, 31 citations in OpenAlex.
- Identification of Shared Regulators of IgE- and MRGPRX2-Dependent Degranulation in Human Mast Cells.Allergy · 2026Article
- Successful Application of the Mast Cell Activation Test in Immediate Hypersensitivity to Amoxicillin.Allergy · 2026Article
- Review
- Mast cell proteases and their significance in physiology, pathology, and therapeutic approaches.Pharmacological reviews · 2026Review
- Mast cell-specific receptor MrgprB2 selectively mediates oxaliplatin-induced neuropathic pain.Journal of translational medicine · 2026Article
- Generation of Functionally Competent Human Mast Cells from Limited Blood Volumes.International journal of molecular sciences · 2026Article
- Evolution of mast cell research associated with the Laboratory of Allergic Diseases, 1985-2022: Summary of the 2023 NIAID Symposium.The Journal of allergy and clinical immunology · 2025Review
- The Anti-Angiogenic Effect of Microbotox on Rosacea Is Due to the Suppressed Secretion of VEGF by Mast Cells Resulting From Internalization of the MRGPRX2 Receptor.Annals of dermatology · 2025Article
- PRL2 negatively regulates FcεRI mediated activation of mast cells.Cell death & disease · 2025Article
- Fedratinib and gandotinib induce apoptosis and enhance the efficacy of tyrosine kinase inhibitors in human mast cells.American journal of cancer research · 2025Article
- LRO biogenesis and function: what can we learn from mast cells?Frontiers in cell and developmental biology · 2025Review
- Agonism of the glutamate receptor GluK2 suppresses dermal mast cell activation and cutaneous inflammation.Science translational medicine · 2024Article
- CRISPR/Cas9-mediated gene disruption determines the roles of MITF and CITED2 in human mast cell differentiation.Blood advances · 2024Article
- Article
- Mechanobiology of Type 1 hypersensitivity: Elucidating the impacts of mechanical forces in allergic reactions.Mechanobiology in medicine · 2024Review
- Insights gained from single-cell analysis of chimeric antigen receptor T-cell immunotherapy in cancer.Military Medical Research · 2023Review
- Siglec-9 is an inhibitory receptor on human mast cells in vitro.The Journal of allergy and clinical immunology · 2023Article
- Distinct Epithelial-Innate Immune Cell Transcriptional Circuits Underlie Airway Hyperresponsiveness in Asthma.American journal of respiratory and critical care medicine · 2023Article
- Human Tumor Targeted Cytotoxic Mast Cells for Cancer Immunotherapy.Frontiers in oncology · 2022Article
- Harnessing the Anti-Tumor Mediators in Mast Cells as a New Strategy for Adoptive Cell Transfer for Cancer.Frontiers in oncology · 2022Review
Corrections and comments
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Authors and funding
7 authors at 4 institutions in 4 countries.
Funding
Abstract
Targeted functional genomics represents a powerful approach for studying gene function in vivo and in vitro. However, its application to gene expression studies in human mast cells has been hampered by low yields of human mast cell cultures and their poor transfection efficiency. We developed an imaging system in which mast cell degranulation can be visualized in single cells subjected to shRNA knockdown or CRISPR-Cas9 gene editing. By using high-resolution confocal microscopy and a fluorochrome-labeled avidin probe, one can directly assess the alteration of functional responses, i.e., degranulation, in single human mast cells (10-12 weeks old). The elimination of a drug or marker selection step avoids the use of potentially toxic treatment procedures, and the brief hands-on time of the functional analysis step enables high-throughput screening of shRNA or CRISPR-Cas9 constructs to identify genes that regulate human mast cell degranulation. The ability to analyze single cells substantially reduces the total number of cells required and enables the parallel visualization of the degranulation profiles of both edited and non-edited mast cells, offering a consistent internal control not found in other protocols. Moreover, our protocol offers a flexible choice between RNA interference (RNAi) and CRISPR-Cas9 genome editing for perturbation of gene expression using our human mast cell single-cell imaging system. Perturbation of gene expression, acquisition of microscopy data and image analysis can be completed within 5 d, requiring only standard laboratory equipment and expertise.
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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.