ArticleAmerican journal of physiology. Lung cellular and molecular physiology2013
Efficient delivery of RNA interference oligonucleotides to polarized airway epithelia in vitro.
Article in American journal of physiology. Lung cellular and molecular physiology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed.
- General trends in the calnexin-dependent expression and pharmacological rescue of clinical CFTR variants.eLife · 2025Article
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- General Trends in the Calnexin-Dependent Expression and Pharmacological Rescue of Clinical CFTR Variants.bioRxiv : the preprint server for biology · 2025Article
- Combinatorial development of nebulized mRNA delivery formulations for the lungs.Nature nanotechnology · 2024Article
- WNK Inhibition Increases Surface Liquid pH and Host Defense in Cystic Fibrosis Airway Epithelia.American journal of respiratory cell and molecular biology · 2022Article
- FXYD3 facilitates NaAmerican journal of physiology. Cell physiology · 2022Article
- Lipid-driven CFTR clustering is impaired in cystic fibrosis and restored by corrector drugs.Journal of cell science · 2022Article
- Analysis of multiple gene co-expression networks to discover interactions favoring CFTR biogenesis and ΔF508-CFTR rescue.BMC medical genomics · 2021Article
- Inflammatory cytokines TNF-α and IL-17 enhance the efficacy of cystic fibrosis transmembrane conductance regulator modulators.The Journal of clinical investigation · 2021Article
- V-Type ATPase Mediates Airway Surface Liquid Acidification in Pig Small Airway Epithelial Cells.American journal of respiratory cell and molecular biology · 2021Article
- Passive siRNA transfection method for gene knockdown in air-liquid interface airway epithelial cell cultures.American journal of physiology. Lung cellular and molecular physiology · 2021Article
- TNFα and IL-17 alkalinize airway surface liquid through CFTR and pendrin.American journal of physiology. Cell physiology · 2020Article
- Article
- Consecutive Hypoxia Decreases Expression ofFrontiers in cell and developmental biology · 2020Article
- Slowing ribosome velocity restores folding and function of mutant CFTR.The Journal of clinical investigation · 2019Article
- Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia.Nature communications · 2019Article
- Design, mechanism, delivery and therapeutics of canonical and Dicer-substrate siRNA.Asian journal of pharmaceutical sciences · 2019Review
- Sialylation of MUC4β N-glycans by ST6GAL1 orchestrates human airway epithelial cell differentiation associated with type-2 inflammation.JCI insight · 2019Article
- Oxidative stress associated with aging activates protein kinase Cε, leading to cilia slowing.American journal of physiology. Lung cellular and molecular physiology · 2018Article
- Article
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Authors and funding
7 authors.
Funding
Abstract
Polarized and pseudostratified primary airway epithelia present barriers that significantly reduce their transfection efficiency and the efficacy of RNA interference oligonucleotides. This creates an impediment in studies of the airway epithelium, diminishing the utility of loss-of-function as a research tool. Here we outline methods to introduce RNAi oligonucleotides into primary human and porcine airway epithelia grown at an air-liquid interface and difficult-to-transfect transformed epithelial cell lines grown on plastic. At the time of plating, we reverse transfect small-interfering RNA (siRNA), Dicer-substrate siRNA, or microRNA oligonucleotides into cells by use of lipid or peptide transfection reagents. Using this approach we achieve significant knockdown in vitro of hypoxanthine-guanine phosphoribosyltransferase, IL-8, and CFTR expression at the mRNA and protein levels in 1-3 days. We also attain significant reduction of secreted IL-8 in polarized primary pig airway epithelia 3 days posttransfection and inhibition of CFTR-mediated Cl⁻ conductance in polarized air-liquid interface cultures of human airway epithelia 2 wk posttransfection. These results highlight an efficient means to deliver RNA interference reagents to airway epithelial cells and achieve significant knockdown of target gene expression and function. The ability to reliably conduct loss-of-function assays in polarized primary airway epithelia offers benefits to research in studies of epithelial cell homeostasis, candidate gene function, gene-based therapeutics, microRNA biology, and targeting the replication of respiratory viruses.
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