Article
Biotechnology & Applied Microbiology
Yifei Wang, Chunxi Wang, Justin Sylvers, Tatiana Segura, Fan Yuan
Summary: Electrotransfection (ET) is a non-viral approach used for delivering nucleic acids, but its efficiency is often low in vivo. In this study, we developed nanoenhancers that diffuse slowly in tissues and are poorly absorbed by blood and lymph microvessels. These nanoenhancers were found to enhance ET efficiency both in vitro and in vivo, and their mechanism of action was related to the inhibition of lysosomal functions.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Matej Kranjc, Janja Dermol-Cerne, Tjasa Potocnik, Vitalij Novickij, Damijan Miklavcic
Summary: This study demonstrates the feasibility of using high-intensity pulsed electromagnetic field (HI-PEMF) for non-viral delivery of genetic material in vitro. The results show that HI-PEMF can cause DNA adsorption to the cell membrane and induce gene electrotransfection. Endocytosis is also identified as a possible mechanism for introducing large molecules into cells by HI-PEMF.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Biochemical Research Methods
Pauline Bregigeon, Charlotte Riviere, Laure Franqueville, Christian Vollaire, Julien Marchalot, Marie Frenea-Robin
Summary: Reversible electroporation is a method that uses pulsed electric fields to increase the permeability of cell membranes for the delivery of molecules, such as anticancer drugs. However, handling steps in electroporation protocols may damage cell spheroids, resulting in non-reproducible electric field conditions. A unique microsystem has been developed to culture, monitor, and electroporate regular spheroids in parallel, ensuring consistent electric field conditions for each spheroid. This microsystem has the potential to improve in vitro assays for preclinical evaluation of electroporation-based treatments.
Article
Biochemical Research Methods
Alinaghi Salari, Sila Appak-Baskoy, Imogen R. Coe, John Abousawan, Costin N. Antonescu, Scott S. H. Tsai, Michael C. Kolios
Summary: This study presents a novel acoustofluidic strategy for efficient intracellular delivery in adherent cells, using strong mechanical oscillations mediated by Lamb waves in a microfluidic channel. The method achieves successful delivery of cargo materials with high efficiency and shows versatility across different cell lines. It also demonstrates the tunability of dosage control and highlights unique features such as high viability, scalability, portability, and ease of operation.
Article
Biochemistry & Molecular Biology
Giedre Silkuniene, Uma M. Mangalanathan, Alessandra Rossi, Peter A. Mollica, Andrei G. Pakhomov, Olga Pakhomova
Summary: The study aimed to identify endogenous proteins that either facilitate or impede the permeabilized state of the cell membrane disrupted by nsEP. Knockouts (KOs) of 316 genes encoding for membrane proteins were generated using a LentiArray CRISPR library in U937 human monocytes expressing Cas9 nuclease. Only two KOs for SCNN1A and CLCA1 genes showed a significant reduction in Yo-Pro-1 uptake, indicating their potential involvement in electropermeabilization lesions or lifespan. However, as many as 39 genes were identified as likely contributors to membrane stability or repair after nsEP.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Biochemistry & Molecular Biology
Yifei Wang, Chun-Chi Chang, Fan Yuan
Summary: Electrotransfection is a non-viral method to deliver nucleic acids into cells. This study determined the minimal copy number of plasmid DNA (pDNA) per cell required for transgene expression and developed a statistical framework to predict pDNA copy number in the nucleus. A novel approach was developed to obtain isolated nuclei with minimal contamination by extranuclear pDNA, enabling accurate measurement of intranuclear pDNA. The data showed that the copy number of pDNA per nucleus depended on cell culture period and pDNA dose for electrotransfection, and were used to improve the understanding of pDNA transport kinetics in cells.
BIOELECTROCHEMISTRY
(2023)
Article
Biochemistry & Molecular Biology
Vitalii Kim, Iurii Semenov, Allen S. Kiester, Mark A. Keppler, Bennett L. Ibey, Joel N. Bixler, Ruben M. L. Colunga Biancatelli, Andrei G. Pakhomov
Summary: Changing the electric field direction can inhibit electroporation and electrostimulation by nanosecond electric pulses. This feature, known as bipolar cancellation, enables selective remote targeting with nanosecond electric pulses and reduces neuromuscular side effects.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Biochemistry & Molecular Biology
Dipeng Zhao, Jian Qin, Jie An, Hao Zhang, Junling Li, Hejie Wang, Rong Du, Yongming He
Summary: Electroporation is a non-viral mediated transfection technique that offers advantages such as being harmless, easy to operate, and less expensive. However, there is a need for further improvement in electroporation efficiency, as there is no universal standard electrotransfection step for different cell types, and inappropriate parameters can result in low transfection efficiency and high cell mortality.
MOLECULAR BIOTECHNOLOGY
(2023)
Article
Chemistry, Physical
Xinxin Hang, Zhaocun Huang, Shiqi He, Zhiying Wang, Zaizai Dong, Lingqian Chang
Summary: A nano-electroporation-DNA tensioner platform is developed to achieve high-speed drug delivery and cellular mechanical force analysis, providing a simple yet powerful tool for clinical drug screening and development.
Review
Chemistry, Multidisciplinary
Jeongsoo Hur, Aram J. Chung
Summary: This review provides an overview of recent advances in microfluidic and nanofluidic intracellular delivery strategies and discusses new opportunities and challenges for clinical applications. Microfluidic and nanofluidic approaches as emerging solutions have shown remarkable potential for addressing this open challenge.
Article
Chemistry, Physical
P. R. Sreedevi, K. Suresh
Summary: Delivery of genetic material is an important tool for analyzing and understanding genetic interventions. Traditional methods rely on viral vectors or xenogenic carrier molecules, which can have carcinogenic, immunogenic, and toxic effects. Cold atmospheric plasma-induced transfection is a new physical method that does not require vectors or carriers. It creates temporary pores on cell membranes for gene delivery. This method has been successfully used in various cell lines and mouse models without adverse effects on cell activity and function.
ADVANCES IN COLLOID AND INTERFACE SCIENCE
(2023)
Article
Biotechnology & Applied Microbiology
Gyeong Won Lee, Byeongyeon Kim, Tae Wook Lee, Sang-Gu Yim, Ajeesh Chandrasekharan, Hyewon Kim, Sungyoung Choi, Seung Yun Yang
Summary: This study presents an electric-pulse-driven nanopore-electroporation (nEP) system for localized intracellular drug delivery in deep tissues. The nEP system achieves better transfection efficiency and high cell recovery rate compared to conventional bulk electroporation. In vivo experiments show that the nanopore needle electrode with a side drug-releasing compartment provides better control over dosage release, time, and location of the model agent. The nEP platform offers a safe and effective method for intracellular delivery in deep tissues.
BIOENGINEERING & TRANSLATIONAL MEDICINE
(2023)
Review
Engineering, Electrical & Electronic
Farzad Forouzandeh, Arpys Arevalo, Ahmed Alfadhel, David A. Borkholder
Summary: This report reviews the progress on peristaltic micropumps and discusses various techniques and important parameters involved in mimicking the physiological transport mechanism. It also evaluates research efforts over the past 30 years and provides suggestions for future development.
SENSORS AND ACTUATORS A-PHYSICAL
(2021)
Article
Pharmacology & Pharmacy
Eivina Radzeviciute-Valciuke, Jovita Gecaite, Augustinas Zelvys, Aukse Zinkeviciene, Rokas Zalneravicius, Veronika Malysko-Ptasinske, Ausra Nemeikaite-Ceniene, Vytautas Kaseta, Natalija German, Jurij Novickij, Almira Ramanaviciene, Julita Kulbacka, Vitalij Novickij
Summary: This study demonstrates that MHz frequency bursts of nanosecond pulses can enhance gene delivery and investigates the potential of gold nanoparticles in this process. The combination of microsecond pulses and gold nanoparticles significantly improves gene delivery, depending on the surface charge and size of the nanoparticles. Gold nanoparticles are ineffective in nanosecond pulse treatment, but MHz pulse treatment remains competitive, with high gene delivery efficiency, low reactive oxygen species generation, preserved cellular viability, and easier operation.
Article
Genetics & Heredity
Lucas dos Santos da Silva, William Borges Domingues, Bruna Fagundes Barreto, Amanda Weege da Silveira Martins, Eduardo Nunes Dellagostin, Eliza Rossi Komninou, Carine D. Corcini, Antonio Sergio Varela Junior, Vinicius Farias Campos
Summary: Electroporation using capillary technique on high-quality bull sperm increases internalization of exogenous DNA without affecting mitochondrial activity, leading to reduced sperm motility. Expression of miRNA-122-5p is upregulated in sperm electroporated at 1500 V, and the presence of exogenous DNA does not impact its expression.
