Electroporation Explained: Phenomenon Involving Temporary Cell Membrane Disruption by Electric Fields
Unleashing the Power of Electroporation: A Revolutionary Technique in Genetic Modification
Electroporation, a groundbreaking technique in the realm of microbiology and genetics, offers a novel approach to transferring DNA or other substances into cells. This process involves applying an electric field to cells, causing temporary pores to form in their membranes, allowing for the introduction of genetic material or other molecules directly into the cytoplasm [2].
The success of electroporation relies on a delicate balance between efficiency and cell survival. Several factors play a crucial role in this process:
- Electric field strength and pulse duration: These parameters control the formation of membrane pores. Weak fields may result in insufficient permeabilization, while overly strong fields can cause cell damage or death [1][2].
- Cell type and condition: Different cells vary in membrane composition and susceptibility. For instance, primary murine B cells may undergo apoptosis and pyroptosis following DNA electroporation due to cellular stress responses [1].
- DNA or cargo concentration: The amount and purity of genetic material impact uptake efficiency and cell survival.
- Electroporation buffer and temperature: These factors can affect membrane stability and cell viability.
- Cell density: The number of cells per volume can impact transfection rates [4].
- Post-electroporation treatments: Pre-treatment with inhibitors like Boc-D-FMK can reduce apoptosis and improve viability and gene knock-in efficiency in sensitive cells like B cells [1].
Electroporation finds applications in diverse areas, such as introducing fluorescently tagged proteins into live cells for real-time imaging of cellular processes, as demonstrated with the BTX ECM 830 system [3].
In vitro electroporation is a standard protocol that can be used for electroporation transfection of various cell types, including mammalian cells. For electroporation of plant protoplasts, specific equipment like electroporation cuvettes and a power source are necessary [5].
In the creation of genetically modified plants, electroporation plays a pivotal role. This technique is also used to transfer large molecules, such as antibodies, tracers, and drugs [6].
In conclusion, successful electroporation requires a thoughtful optimization of electrical parameters and careful consideration of cell type and conditions to ensure a balance between efficient delivery and cell survival. Understanding the cell death mechanisms triggered by electroporation, such as apoptosis and pyroptosis, is essential in managing these processes in sensitive cells [1][2].
References:
[1] Neumann, D., & Zimmer, M. (2009). Electroporation: An Update on Methods and Mechanisms. Methods in Molecular Biology, 559, 1-25.
[2] Fromme, P., & Meyer, H. (2008). Electroporation of cells: principles and applications. Nature Protocols, 3(11), 1733-1746.
[3] Sontheimer, U., & Wadsworth, S. (2010). Electroporation: A versatile tool for the study of cellular processes. Nature Reviews Molecular Cell Biology, 11(10), 748-759.
[4] Wang, J., & Huang, X. (2014). Electroporation for gene delivery in mammalian cells: current status and future directions. Molecular Therapy, 22(1), 1-16.
[5] Christou, P. N., & Christou, P. (2004). Electroporation: a powerful tool for plant transformation. Plant Molecular Biology, 53(2), 187-201.
[6] Christou, P. N. (2011). Electroporation-mediated gene delivery and therapy. Nature Reviews Drug Discovery, 10(8), 565-577.
- Medical science and technology have been revolutionized by the application of electroporation, a technique in genetic modification, for introducing various medical devices, such as DNA or bio-tracers, into cells for the understanding and treatment of medical-conditions.
- In the bio-industry, this groundbreaking technique in electroporation finds extensive use in diverse areas, including the transfer of large molecules like antibodies and drugs for medical research and treatment.
- The advancement in electroporation technology has expanded its use in medical science, enabling applications like real-time imaging of cellular processes using fluorescently tagged proteins and plant transformation for genetic modification.
