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Electroporation and Nucleofector® Technology
The ability to introduce DNA, RNA or proteins into cells to alter their genotype or phenotype (a process called transfection) is crucial in a variety of life science applications. Various types of transfection methods exist and choosing which approach to use often depends on its suitability to the application in question. Electroporation is a physical transfection method that permeabilizes the cell membrane by applying an electrical pulse and moves molecules via the electrical field into the cell. It is a powerful tool for transfecting large DNA fragments and achieving good transfection efficiencies in cell lines. However, due to high toxicity traditional electroporation has been less successful for efficiently transfecting more biologically relevant primary cells and stem cells, which has limited its application.
Our solution is an improved electroporation technology, the Nucleofector® Technology, originally introduced into the market by legacy Amaxa in 2001. It enables highly efficient, transfection of primary cells, stem cells, neurons, and cell lines that have traditionally been difficult to transfect via electroporation and other non-viral transfection methods. In recent years, this has opened novel opportunities for disease research and therapeutic development, including the advancement of gene therapies, immunotherapies, and stem cell generation.
In the following sections, we provide an overview of Nucleofector® Technology, and explore what benefits it can bring to your research over traditional electroporation methods. We also summarize the transfection capabilities and flexible scaling offered by our various Nucleofector® Platforms, to help you choose which one is best suited to your specific application. This can ensure you obtain the very highest levels of transfection efficiency and quality for your research.
Nucleofection® Technology Animation
See how transfection with Nucleofection® Technology is different from conventional electroporation. It allows for highly efficient transfer of various substrates directly into the nucleus of cells. The modular 4D-Nucleofector® System helps you scale up from low to high-volume transfection or higher throughput without having to re-optimize.
What is Nucleofector® Technology?
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The Nucleofector® Technology uses a specific combination of optimized electrical parameters and cell type-specific solutions which enables transfer of a molecule directly into the cells’ nucleus. Due to this independency from cell proliferation, it allows for efficient transfection of even non-dividing primary cells (like resting T cells or neurons) and also speeds up expression.
Three specialized components are key to the successful and efficient transfection of primary cells or cell lines using a variety of substrates:
- A Nucleofector® System that comprises unique electrical parameters pre-programmed for each optimized cell type, to deliver the substrate directly into the cell nucleus and the cytoplasm. The different platforms we offer provide different specifications for various applications.
- Nucleofector® Kits, containing dedicated Nucleofector® Solutions and Supplements. These act as a protective environment for high transfection efficiency and cell viability, while maintaining physiologically relevant cellular conditions. Specified Nucleofection vessels, pipettes, and a fluorescent positive control vector (pmaxGFPTM Control Vector) are also provided.
- Optimized protocols offering comprehensive guidance for optimal Nucleofection® Conditions along with tips for cell sourcing, passage, growth conditions and media, and post-transfection culture. You can also learn more about how to optimize Nucleofection® Conditions here.
Since launching the Nucleofector® Technology by legacy Amaxa, we have introduced several innovations around those three components to expand its application to further cell types, lower and higher cell numbers or different throughputs.
What Key Benefits Does Nucleofector® Technology Offer?
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The Nucleofector® Technology offers a higher transfection efficiency than other non-viral transfection methods (including traditional electroporation1) along with other significant advantages outlined below.
With its superior transfection performance, Nucleofection offers various advantages over traditional electroporation methods:
- High transfection efficiencies of up to 90% for plasmid DNA and 99% for oligonucleotides, like siRNA.
- Excellent preservation of the physiological status and viability of transfected cells.
- Analysis of transfection results already shortly after transfection possible.
- Easy to use technology, with over 650 cell-type specific protocols that have led to direct transfection success.
- Transfection of a wide range of substrates, including DNA, mRNA, miRNA, siRNA, peptides or proteins.
- Transfection of hard-to-transfect cells, including primary cells, stem cells, neurons and cell lines, as well as cells in adherence.
Additionally, using Nucleofector® Technology can bring various practical benefits to your research, including:
- Reliance on a proven, reliable and innovative technology, that has featured in more than 8000 peer-reviewed publications worldwide
- Reduced risk of cross-contamination using the disposable sterile Nucleocuvette® Vessels
- Excellent technical and applicative support from a highly-skilled, specialist scientific support team
- Easy expansion of your research due to the flexible scaling capabilities of the different Nucleofector® Platforms, including low-, medium- and high-throughput transfection and easy transference of conditions between different device platforms.
These benefits can facilitate numerous applications, such as therapeutic gene knock-down via RNAi2 or CRISPR3 and the generation of induced pluripotent stem cells4 or CAR-T5, among many others. It is therefore not surprising that Nucleofector® Technology is now used in many different lines of research, including functional and structural genomics, drug discovery, and gene and cell therapy.
1 Maasho et al 2004 Journal of Immunological Methods
2 Marques and Williams, 2005 Nature Biotechnology
3 Seki A, Rutz S, 2018 J Exp Med
4 Kaji et al. 2009 Nature
5 Monjezi et al 2017
Flexible Scaling and Adherent Electroporation via Nucleofection
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4D-Nucleofector® X Unit: Transfecting Various Cell Numbers in Different Formats
The 4D-Nucleofector® X Unit features positions for both 20 µL Nucleocuvette® Strips and 100 µL single Nucleocuvette® Vessels. Conditions can be seamlessly transferred between these two vessel types. Furthermore, cells that have been transfected using the X Unit, can be transferred to other Nucleofector Platforms (96-well Shuttle® Device, HT Nucleofector® System, and 4D-Nucleofector® LV Unit) without the need for re-optimization. Learn more by visiting our 4D-Nucleofector® X Unit product page or by watching the video.
4D-Nucleofector® Y Unit: Adherent Electroporation
Electroporation usually requires cells to be in suspension for transfection. However, the 4D-Nucleofector® Y Unit offers the opportunity to keep cells in adherence during electroporation. Adherent primary cells, especially neurons, at defined developmental stages can be transfected using the Y Unit without affecting their functionality and with an efficiency of up to 70%. To find out more, watch the video or read more information on the 4D-Nucleofector® Y Unit product page.
4D-Nucleofector® 96-well Unit: An Add-On for 96-Well Transfection
The 4D-Nucleofector 96-well® Unit offers a medium-throughput add-on for the 4D-Nucleofector® System. It is suited for the convenient optimization of Nucleofection® Conditions or as an assay establishment tool. The 4D-Nucleofector® 96-well Unit serves as a plate handler that mediates the transfer of the a Nucleofector® Program generated by the 4D-Nucleofector® Core and X Unit to a specific well of the 96-well Nucleocuvette® Plate. For scalable throughput, each well of the plate can be addressed and transfected individually. This is useful for optimizing difficult-to-transfect cell lines in just one plate, as well as transfecting variable cell numbers from 104 – 106 cells per reaction. Learn more by visiting the 4D-Nucleofector® 96-well Unit page here and watching the video tutorial.
4D-Nucleofector® LV Unit: Large-Scale Transfection
The 4D-Nucleofector® LV Unit enables closed, scalable and sterile transfection of larger cell numbers in the range of 1×107 to 1×109 cells. It offers two large-scale transfection formats, 1 mL Nucleocuvette® Cartridges or LV Nucleocuvette® Cartridges for a processing volume of up to 20 mL, depending on the application and how many cells need to be transfected. The large-scale transfection capabilities of the LV Unit have made it suitable for a wide range of applications. GMP grade TheraPEAK® Consumables, and a 21CFR part 11 compliant software and IQOQ services allows you to quickly translate research results into a GMP compliant cell therapy manufacturing process. You can read our 4D-Nucleofector® LV Unit product page for more information.
HT Nucleofector® System
High Throughput Transfection
The HT Nucleofector® System is an independent platform offering high-throughput transfection in 384-well format. With an extremely fast plate processing time of one minute and high reproducibility it is the ideal tool for screening applications. Find out more by visiting the HT Nucleofector® System product page or watching our video tutorial.
