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Non-Viral Delivery of Genome-Editing Nucleases for Gene Therapy
Published in Yashwant Pathak, Gene Delivery, 2022
Electroporation is known as a physical transfection method in which high voltage current is applied to the cells to enhance the membrane permeability to nucleic acids, chemicals, or proteins. Due to application of the electrical impulse, small gaps between the cell membranes appear that are resealable allowing the entry of substances [3]. Initial in vitro studies proved that the electroporation method delivered the plasmid DNA containing Cas9 efficiently to the cells which are considered difficult to transfect, such as primary fibroblasts [3, 11], human embryonic stem cells [3, 11], pluripotent stem cells , and neurons. However, therapeutic application of electroporation mediated-delivery is only limited to ex vivo. Currently, clinical trials by this method are taking place [Table 12.2]. Many groups have utilized electroporation to directly deliver nucleic acids or proteins for ZFNs, TALENs, or CRISPR ribonucleoproteins, and many ongoing clinical trials are utilizing electroporation for delivery to T cells. [3, 11–13]
Combination of Microneedles with Other Methods
Published in Boris Stoeber, Raja K Sivamani, Howard I. Maibach, Microneedling in Clinical Practice, 2020
Delivery of FITC-Dextran with a molecular weight of 4.3 kDa was evaluated in vitro by Yan et al. Skin was pretreated with microneedles followed by application of high-voltage pulses using electrodes. Two types of electroporation were tested: in-skin and on-skin. Delivery was found to be higher with the combination approach for both the electroporation methods when compared to microneedles or electroporation alone. Delivery was found to be dependent on voltage and pulse width. Permeability with on-skin electroporation was 20-fold higher and with in-skin electroporation it was 140-fold higher. No significant skin irritation was observed (36).
Radio-Electro-Chemotherapy of Cancer: New Perspectives for Cancer Treatment
Published in Pandit B. Vidyasagar, Sagar S. Jagtap, Omprakash Yemul, Radiation in Medicine and Biology, 2017
Pratip Shil, Pandit B. Vidyasagar, Kaushala Prasad Mishra
Electroporation involves the enhancement of the plasma membrane permeability by exposing the biological cells to electric pulses of high voltage and short durations. As early as 1967, Sale and Hamilton [14] had observed the killing of microorganisms, viz., bacteria and yeast due to exposure to high-voltage electric pulses. It was observed in 1972 by Neumann and Rosenheck [15] that the exposure to electric field generated changes in the permeability of the biological cells. This was followed by the study of electric field induced pore formation in plasma membrane by Kinosita and Tsong [16].
Microneedle technology for potential SARS-CoV-2 vaccine delivery
Published in Expert Opinion on Drug Delivery, 2023
Megan McNamee, Shuyi Wong, Owen Guy, Sanjiv Sharma
As a result of increased funding following the SARS-CoV-2 pandemic, new medical vaccination devices are rapidly emerging. Exciting work has been published by the Georgia Institute of Technology, with data published evidencing a well-tolerated, low-cost MN electrode ePatch [133]. This portable device utilizes a piezoelectric pulse that, following thumb pressure, enables an MN array with dense electrode spacing to create and deliver high electric field pulses to the epidermis [133]. With traditional electroporation widely reported to leave enduring damage to the epidermis, the short length of the MNA prevented significant damage to the epidermal integrity of rat and murine skin [133]. Promising data has been reported following the use of the ePatch, with the delivery of the S protein inducing robust immunogenicity and specific antibody response, demonstrating a dose-sparing potential of 10-fold when compared to conventional intramuscular and intradermal DNA vaccination [133]. Further studies are required, however, to demonstrate the potential impact on the integrity of human tissue following electroporation with an MN patch and to perform investigations into pain ranking to ensure that combinations of this technology offer the same nociception benefits as previously reported.
Progress of delivery methods for CRISPR-Cas9
Published in Expert Opinion on Drug Delivery, 2022
Wu Yang, Jiaqi Yan, Pengzhen Zhuang, Tao Ding, Yu Chen, Yu Zhang, Hongbo Zhang, Wenguo Cui
Electroporation is another common physical method. It delivers hydrodynamic nanometers in diameter component nucleic acids or proteins into cells by opening the phospholipid bilayer on the cell membrane of cells suspended in buffer using pulsed, high voltage current(Figure 3B) [43]. Electroporation has been applied for in vivo and in vitro delivery of the CRISPR-Cas9 system in mammals [44,45]. However, electroporation is usually not suitable for in vivo applications due to a large amount of voltage that usually needs to be applied across the cell membrane, which can cause damage to the phospholipid bilayer and ultimately lead to apoptosis or poor cell activity. To minimize the side effects caused by electroporation, the researchers optimized the voltage, current parameters, and medium composition for this method [46]. Due to the appeal of exploiting the often-used laboratory research and efficient CRISPR-Cas9 conversion, electroporation will likely continue to be used and improved as the primary technique for efficient delivery of the CRISPR-Cas9 system.
Response characteristics and optimization of electroporation: simulation based on finite element method
Published in Electromagnetic Biology and Medicine, 2021
Cheng Zhou, Zeyao Yan, Kefu Liu
Electroporation involves exposing cells to external high-voltage electric pulses that cause an increase of the transmembrane voltage (TMV), rearrange membrane phospholipids, and create pores in the cell membrane (Weaver and Chizmadzhev 1996). By increasing membrane permeability, molecules, and ions that are normally prevented from crossing the membrane are now able to pass into or out of cells. Since its inception in the 1980s, electroporation has been used for gene transfer (Weaver and Chizmadzhev 1996) (Mella et al. 2019) and therapy (Neumann et al. 1982), gene editing (Laustsen and Bak 2019), transdermal drug delivery (Prausnitz et al. 1993), tumor ablation (Al-Sakere et al. 2007), cell fusion (Jordan et al. 2013), extraction of useful compounds from plants (Sack et al. 2010) (Loginova et al. 2011), and sterilization of milk and juice (Food and Drug Administration 2000) (Sato et al. 2019).