ArticleBioelectrochemistry (Amsterdam, Netherlands)2021
A statistical framework for determination of minimal plasmid copy number required for transgene expression in mammalian cells.
Article in Bioelectrochemistry (Amsterdam, Netherlands), 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 14 citations in OpenAlex.
- Quantitative Analysis of Plasmid DNA Distribution and Transgene Expression Following Cell Squeezing.Annals of biomedical engineering · 2026Article
- Stepwise Evaluation of Plasmid- and Adeno-Associated Virus-Based Knock-In Using A Triple-Reporter Platform.Genes to cells : devoted to molecular & cellular mechanisms · 2026Article
- Mechanisms and Strategies for Enhancing DNA Nuclear Entry in Gene Delivery.Cellular and molecular bioengineering · 2026Article
- Electroporation- and Liposome-Mediated Co-Transfection of Single and Multiple Plasmids.Pharmaceutics · 2025Article
- Perfusion-Based Production of rAAV via an Intensified Transient Transfection Process.Biotechnology and bioengineering · 2025Article
- Copy number of naked DNA delivered into nucleus of mammalian cells by electrotransfection.Bioelectrochemistry (Amsterdam, Netherlands) · 2023Article
- Nanoenhancer for improving naked DNA electrotransfectionFrontiers in bioengineering and biotechnology · 2023Article
- Effect of Experimental Electrical and Biological Parameters on Gene Transfer by Electroporation: A Systematic Review and Meta-Analysis.Pharmaceutics · 2022Review
- Gene Electrotransfer into Mammalian Cells Using Commercial Cell Culture Inserts with Porous Substrate.Pharmaceutics · 2022Article
- Article
- Non-Clinical In Vitro Evaluation of Antibiotic Resistance Gene-Free Plasmids Encoding Human or Murine IL-12 Intended for First-in-Human Clinical Study.Pharmaceutics · 2021Article
- Evaluation of a Novel Plasmid for Simultaneous Gene Electrotransfer-Mediated Silencing of CD105 and CD146 in Combination with Irradiation.International journal of molecular sciences · 2021Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Plasmid DNA (pDNA) has been widely used for non-viral gene delivery. After pDNA molecules enter a mammalian cell, they may be trapped in subcellular structures or degraded by nucleases. Only a fraction of them can function as templates for transcription in the nucleus. Thus, an important question is, what is the minimal amount of pDNA molecules that need to be delivered into a cell for transgene expression? At present, it is technically a challenge to experimentally answer the question. To this end, we developed a statistical framework to establish the relationship between two experimentally quantifiable factors - average copy number of pDNA per cell among a group of cells after transfection and percent of the cells with transgene expression. The framework was applied to the analysis of electrotransfection under different experimental conditions in vitro. We experimentally varied the average copy number per cell and the electrotransfection efficiency through changes in extracellular pDNA dose, electric field strength, and pulse number. The experimental data could be explained or predicted quantitatively by the statistical framework. Based on the data and the framework, we could predict that the minimal number of pDNA molecules in the nucleus for transgene expression was on the order of 10. Although the prediction was dependent on the cell and experimental conditions used in the study, the framework may be generally applied to analysis of non-viral gene delivery.
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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.