Effect of Experimental Electrical and Biological Parameters on Gene Transfer by Electroporation: A Systematic Review and Meta-Analysis

Potočnik, Tjaša; Lebar, Alenka Maček; Kos, Špela; Reberšek, Matej; Pirc, Eva; Serša, Gregor; Miklavčič, Damijan

doi:10.3390/pharmaceutics14122700

Cited by 5 publications

(4 citation statements)

References 265 publications

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“…Furthermore, using the same pulses (either long or short, or bipolar) for delivering cytotoxic drugs into the cells, as well as pDNA to achieve simultaneous gene electrotransfer, is an attractive idea that may be within reach. 94 …”

Section: Discussionmentioning

confidence: 99%

The equivalence of different types of electric pulses for electrochemotherapy with cisplatin − an in vitro study

Scuderi,

Dermol-Cerne,

Scancar

et al. 2024

Radiology and Oncology

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Background Electrochemotherapy (ECT) is a treatment involving the administration of chemotherapeutics drugs followed by the application of 8 square monopolar pulses of 100 μs duration at a repetition frequency of 1 Hz or 5000 Hz. However, there is increasing interest in using alternative types of pulses for ECT. The use of high-frequency short bipolar pulses has been shown to mitigate pain and muscle contractions. Conversely, the use of millisecond pulses is interesting when combining ECT with gene electrotransfer for the uptake of DNA-encoding proteins that stimulate the immune response with the aim of converting ECT from a local to systemic treatment. Therefore, the aim of this study was to investigate how alternative types of pulses affect the efficiency of the ECT. Materials and methods We performed in vitro experiments, exposing Chinese hamster ovary (CHO) cells to conventional ECT pulses, high-frequency bipolar pulses, and millisecond pulses in the presence of different concentrations of cisplatin. We determined cisplatin uptake by inductively coupled plasma mass spectrometry and cisplatin cytotoxicity by the clonogenic assay. Results We observed that the three tested types of pulses potentiate the uptake and cytotoxicity of cisplatin in an equivalent manner, provided that the electric field is properly adjusted for each pulse type. Furthermore, we quantified that the number of cisplatin molecules, resulting in the eradication of most cells, was 2−7 × 107 per cell. Conclusions High-frequency bipolar pulses and millisecond pulses can potentially be used in ECT to reduce pain and muscle contraction and increase the effect of the immune response in combination with gene electrotransfer, respectively.

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Section: Discussionmentioning

confidence: 99%

The equivalence of different types of electric pulses for electrochemotherapy with cisplatin − an in vitro study

Scuderi,

Dermol-Cerne,

Scancar

et al. 2024

Radiology and Oncology

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“…During macroscale electroporation, also called bulk or cuvette electroporation, multiple cells are treated at once in chambers with a diameter of at least 1 mm, providing a straightforward, inexpensive, and high-throughput transfection method [ 184 ].…”

Section: Intracellular Deliverymentioning

confidence: 99%

Basic Principles of RNA Interference: Nucleic Acid Types and In Vitro Intracellular Delivery Methods

Isenmann,

Stoddart,

Schmelzeisen

et al. 2023

Micromachines

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Since its discovery in 1989, RNA interference (RNAi) has become a widely used tool for the in vitro downregulation of specific gene expression in molecular biological research. This basically involves a complementary RNA that binds a target sequence to affect its transcription or translation process. Currently, various small RNAs, such as small interfering RNA (siRNA), micro RNA (miRNA), small hairpin RNA (shRNA), and PIWI interacting RNA (piRNA), are available for application on in vitro cell culture, to regulate the cells’ gene expression by mimicking the endogenous RNAi-machinery. In addition, several biochemical, physical, and viral methods have been established to deliver these RNAs into the cell or nucleus. Since each RNA and each delivery method entail different off-target effects, limitations, and compatibilities, it is crucial to understand their basic mode of action. This review is intended to provide an overview of different nucleic acids and delivery methods for planning, interpreting, and troubleshooting of RNAi experiments.

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“…Electroporation (under this name) was first described 50 years ago ( Neumann and Rosenheck, 1972 ) and causes transient increase in permeability of the cell membrane by applying high-voltage electric field pulses. This technique has gained attention as an effective, non-chemical method for introducing molecules into cells, including foreign DNA ( Potocnik et al, 2022 ; Rakoczy et al, 2022 ) and membrane-impermeable anticancer drugs (i.e., electrochemotherapy) ( Yarmush et al, 2014 ). At stronger electric fields, cells are damaged, leading to cell death.…”

Section: Introductionmentioning

confidence: 99%

Inactivation of antibiotic-resistant bacteria Escherichia coli by electroporation

Haberl Meglič,

Slokar,

Miklavčič

2024

Front. Microbiol.

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IntroductionIn modern times, bacterial infections have become a growing problem in the medical community due to the emergence of antibiotic-resistant bacteria. In fact, the overuse and improper disposal of antibiotics have led to bacterial resistance and the presence of such bacteria in wastewater. Therefore, it is critical to develop effective strategies for dealing with antibiotic-resistant bacteria in wastewater. Electroporation has been found to be one of the most promising complementary techniques for bacterial inactivation because it is effective against a wide range of bacteria, is non-chemical and is highly optimizable. Many studies have demonstrated electroporation-assisted inactivation of bacteria, but rarely have clinical antibiotics or bacteria resistant to these antibiotics been used in the study. Therefore, the motivation for our study was to use a treatment regimen that combines antibiotics and electroporation to inactivate antibiotic-resistant bacteria.MethodsWe separately combined two antibiotics (tetracycline and chloramphenicol) to which the bacteria are resistant (with a different resistance mode) and electric pulses. We used three different concentrations of antibiotics (40, 80 and 150 µg/ml for tetracycline and 100, 500 and 2000 µg/ml for chloramphenicol, respectively) and four different electric field strengths (5, 10, 15 and 20 kV/cm) for electroporation.Results and discussionOur results show that electroporation effectively enhances the effect of antibiotics and inactivates antibiotic-resistant bacteria. The inactivation rate for tetracycline or chloramphenicol was found to be different and to increase with the strength of the pulsed electric field and/or the concentration of the antibiotic. In addition, we show that electroporation has a longer lasting effect (up to 24 hours), making bacteria vulnerable for a considerable time. The present work provides new insights into the use of electroporation to inactivate antibiotic-resistant bacteria in the aquatic environment.

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Effect of Experimental Electrical and Biological Parameters on Gene Transfer by Electroporation: A Systematic Review and Meta-Analysis

Cited by 5 publications

References 265 publications

The equivalence of different types of electric pulses for electrochemotherapy with cisplatin − an in vitro study

The equivalence of different types of electric pulses for electrochemotherapy with cisplatin − an in vitro study

Basic Principles of RNA Interference: Nucleic Acid Types and In Vitro Intracellular Delivery Methods

Inactivation of antibiotic-resistant bacteria Escherichia coli by electroporation

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