ArticleFrontiers in physiology2016
A Guide to Transient Expression of Membrane Proteins in HEK-293 Cells for Functional Characterization.
Article in Frontiers in physiology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 46 papers, 1 of them a synthesis that pooled it.
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Who cites it
46 citing papers in PubMed, 1 synthesis or guideline pooled it, 88 citations in OpenAlex.
- Molecular determinants for brain targeting by peptides: a meta-analysis approach with experimental validation.Fluids and barriers of the CNS · 2024Pooled it
- Direct targeting of ORAI1 by ginsenoside Rg3 modulates calcium signaling and senescence-associated AMPK-NRF2 activation.Journal of ginseng research · 2026Article
- Glucose transporter 1 deficiency impairs glucose homeostasis, cell proliferation, and morphology in human embryonic kidney cells 293.Cell and tissue research · 2026Article
- AMPK-mediated HCN4 channel phosphorylation contributes to age-related intrinsic bradycardia.The Journal of general physiology · 2026Article
- Aquaporin-1, aquaporin-3 and aquaporin-5 differentially modulate cell biophysical and biomechanical properties, impacting cell stiffness and cell-cell adhesion.The FEBS journal · 2026Article
- Integrated Modular Workflows for Membrane Protein Production in HEK293 Cells.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Diethylcarbamazine elicits calcium signals by activation of Brugia malayi TRP-2b channels heterologously expressed in HEK293 cells.Scientific reports · 2025Article
- Long QT syndrome type 1: clinical and functional characterization of KCNQ1 variant c.1111G > C.BMC cardiovascular disorders · 2025Article
- Diethylcarbamazine elicits calcium signals by activation of Brugia malayi TRP-2b channels heterologously expressed in HEK293 cells.Research square · 2025Article
- Identification and characterisation of subtype-specific anti-N-CoR OSGEP protease in acute myeloid leukaemia (AML-M5) cell lineage.Biotechnology letters · 2025Article
- CRISPR-Cas9-Mediated ATF6B Gene Editing Enhances Membrane Protein Production in HEK293T Cells.Bioengineering (Basel, Switzerland) · 2025Article
- Investigation of enhanced adhesion of HEK293T cells on SAM-modified ITO surfaces using NMR metabolomics.Frontiers in bioengineering and biotechnology · 2025Article
- Predicting oxytocin binding dynamics in receptor genetic variants through computational modeling.npj women's health · 2025Article
- Factors affecting rAAV titers during triple-plasmid transient transfection in HEK-293 cells.Biotechnology letters · 2024Article
- Discovery of Therapeutic Antibodies Targeting Complex Multi-Spanning Membrane Proteins.BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy · 2024Review
- Prolonged Activation of the GLP-1 Receptor via Covalent Capture.ACS chemical biology · 2024Article
- In vitro human cell culture models in a bench-to-bedside approach to epilepsy.Epilepsia open · 2024Review
- Cutting-Edge HEK293T Protein-Integrated Lipid Nanostructures: Boosting Biocompatibility and Efficacy.International journal of molecular sciences · 2024Article
- Chitosan functionalized MnCommunications biology · 2023Article
- Passive Recording of Bioelectrical Signals from Non-Excitable Cells by Fluorescent Mirroring.Nano letters · 2023Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The human embryonic kidney 293 (HEK-293) cells are commonly used as host for the heterologous expression of membrane proteins not least because they have a high transfection efficiency and faithfully translate and process proteins. In addition, their cell size, morphology and division rate, and low expression of native channels are traits that are particularly attractive for current-voltage measurements. Nevertheless, the heterologous expression of complex membrane proteins such as receptors and ion channels for biological characterization and in particular for single-cell applications such as electrophysiology remains a challenge. Expression of functional proteins depends largely on careful step-by-step optimization that includes the design of expression vectors with suitable identification tags, as well as the selection of transfection methods and detection parameters appropriate for the application. Here, we use the heterologous expression of a plant potassium channel, the Arabidopsis thaliana guard cell outward-rectifying K(+) channel, AtGORK (At5G37500) in HEK-293 cells as an example, to evaluate commonly used transfection reagents and fluorescent detection methods, and provide a detailed methodology for optimized transient transfection and expression of membrane proteins for in vivo studies in general and for single-cell applications in particular. This optimized protocol will facilitate the physiological and cellular characterization of complex membrane proteins.
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