Predicting electrotransfer in ultra-high frequency sub-microsecond square wave electric fields

Murauskas, Arūnas; Staigvila, Gediminas; Girkontaitė, Irutė; Zinkevičienė, Auksė; Ruzgys, Paulius; Šatkauskas, Saulius; Novickij, Jurij; Novickij, Vitalij

doi:10.1080/15368378.2019.1710529

Cited by 9 publications

(3 citation statements)

References 35 publications

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“…However, for nanosecond pulses, higher frequencies increase efficiency [ 179 ]. Very high frequency pulses can accumulate in cells and reduce the threshold of energy required for RE [ 180 , 181 ]. In contrast, very low frequencies (0.1–1 Hz) can increase efficiency by electrosensitization of the membrane [ 182 , 183 ].…”

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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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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show abstract

“…Nevertheless, a further increase in repetition frequency of nanosecond pulses triggers a new high-frequency (500 kHz–1 MHz) phenomenon of transmembrane potential accumulation resulting in a significantly increased electroporation efficiency ( 121 , 125 , 127 ). It was shown that it is possible to achieve a threshold repetition frequency when the discharging [transmembrane potential (TMP) relaxation] time of the membrane is higher than the delay between the pulses; thus, the TMP starts to accumulate throughout the burst ( 121 , 128 ). The phenomenon in some studies is referred to as “MHz pulse compression” ( 129 ) and allows one to significantly lower the electroporation threshold using the pulse repetition frequency modulation.…”

Section: The Effects Of Pulse Repetition Ratementioning

confidence: 99%

Does the shape of the electric pulse matter in electroporation?

et al. 2022

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Electric pulses are widely used in biology, medicine, industry, and food processing. Numerous studies indicate that electroporation (EP) is a pulse-dependent process, and the electric pulse shape and duration strongly determine permeabilization efficacy. EP protocols are precisely planned in terms of the size and charge of the molecules, which will be delivered to the cell. In reversible and irreversible EP applications, rectangular or sine, polar or bipolar pulses are commonly used. The usage of pulses of the asymmetric shape is still limited to high voltage and low voltage (HV/LV) sequences in the context of gene delivery, while EP-based applications of ultra-short asymmetric pulses are just starting to emerge. This review emphasizes the importance and role of the pulse shape for membrane permeabilization by EP.

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“…Nevertheless, the proof of concept was shown recently [13,44], and as expected, ultrashort pulses are less efficient for gene delivery, however, only when delivered with several Hz or kHz frequencies. In our previous studies, we have shown that compressing nanosecond pulses into an MHz burst triggers a new phenomenon of residual TMP accumulation, which significantly improves electrotransfer [45,46] and, thus, can be applied in nanosecond range electrogene delivery context [14].…”

Section: Introductionmentioning

confidence: 99%

Improving NonViral Gene Delivery Using MHz Bursts of Nanosecond Pulses and Gold Nanoparticles for Electric Field Amplification

et al. 2023

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Gene delivery by the pulsed electric field is a promising alternative technology for nonviral transfection; however, the application of short pulses (i.e., nanosecond) is extremely limited. In this work, we aimed to show the capability to improve gene delivery using MHz frequency bursts of nanosecond pulses and characterize the potential use of gold nanoparticles (AuNPs: 9, 13, 14, and 22 nm) in this context. We have used bursts of MHz pulses 3/5/7 kV/cm × 300 ns × 100 and compared the efficacy of the parametric protocols to conventional microsecond protocols (100 µs × 8, 1 Hz) separately and in combination with nanoparticles. Furthermore, the effects of pulses and AuNPs on the generation of reactive oxygen species (ROS) were analyzed. It was shown that gene delivery using microsecond protocols could be significantly improved with AuNPs; however, the efficacy is strongly dependent on the surface charge of AuNPs and their size. The capability of local field amplification using AuNPs was also confirmed by finite element method simulation. Finally, it was shown that AuNPs are not effective with nanosecond protocols. However, MHz protocols are still competitive in the context of gene delivery, resulting in low ROS generation, preserved viability, and easier procedure to trigger comparable efficacy.

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Predicting electrotransfer in ultra-high frequency sub-microsecond square wave electric fields

Cited by 9 publications

References 35 publications

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

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

Does the shape of the electric pulse matter in electroporation?

Improving NonViral Gene Delivery Using MHz Bursts of Nanosecond Pulses and Gold Nanoparticles for Electric Field Amplification

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