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What is Nucleofector® Technology?

The Nucleofector® Technology, introduced to the market by legacy Amaxa in 2001, uses a specific combination of optimized electrical parameters and cell type-specific solutions, which enables transfer of a molecule directly into the cell’s nucleus. Thus, Nucleofection® is independent from cell proliferation and allows the transfection of non-dividing primary cells (e.g. resting T cells or neurons). 
Three specialized components are key to the efficient transfection of primary cells or cell lines using a variety of substrates:
  1.  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. Find further information below.
  2. 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.
  3. 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.
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Nucleofector® Technology – Key Benefits

  • Transfection of a wide range of substrates, including DNA, mRNA, RNPs, miRNA, siRNA, peptides or proteins
  • Higher transfection efficiency than other non-viral transfection methods (including traditional electroporation1)
  • 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 shortly after transfection possible Easy-to-use technology, with over 750 cell-type specific protocols that have led to direct transfection success
  • 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 21,500 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 transfer of conditions between different platforms (find further information below)
     

These benefits can facilitate numerous applications, such as therapeutic gene knock-down via RNAi2, genome editing via CRISPR3,4, the generation and transfection of induced pluripotent stem cells5,6 or generation of CAR-T cells7,8, among many others. It is therefore not surprising that Nucleofector® Technology is used in many different lines of research, including functional and structural genomics, drug discovery, and gene and cell therapy.

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Nucleofector® Platforms – What is the Right System for You?

We have developed different Nucleofector® Platforms that offer a range of different specifications and units to enable flexible scaling (i.e., Nucleofection® of varying numbers and volume of cells depending on the application) as well as transfection of adherent cells.

4D-Nucleofector® System

This advanced, flexible and convenient 4D-Nucleofector® System comprises a Core Unit, as well as several functional units with different transfection capabilities to suit different applications. Find further information below or watch this video.

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4D-Nucleofector® Platform - The Story Of

See how breakthrough Nucleofector® Technology innovations in transfection and electroporation culminate in the next generation 4D-Nucleofector® System.

4D-Nucleofector® Core Unit: The Controlling Unit 
 


The 4D-Nucleofector® Core Unit is the heart of the 4D-Nucleofector® System and needed to control the functional units. As the 4D-Nucleofecor® Core Unit can control several functional units you can set up your 4D-Nucleofector® System depending on your needs. An intuitive, touch-screen-based user interface allows you to design and save experiments. In addition, the 4D-Nucleofector® Core Unit comes with more than 160 predefined Nucleofection® Parameters for cell lines and primary cells.

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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 (4D-Nucleofector® 96-well Unit, 384-well Nucleofector® System, and 4D-Nucleofector® LV Unit) without the need for re-optimization. Learn more by visiting our 4D-Nucleofector® X Unit product page.

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4D-Nucleofector® Y Unit: Adherent Electroporation 
 


Electroporation usually requires cells to be in suspension for transfection. 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 4D-Nucleofector® Y Unit without affecting their functionality and with an efficiency of up to 70%. To find out more, visit the 4D-Nucleofector® Y Unit product page.

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4D-Nucleofector® 96-well Unit: Increased Throughput – 96-Well Transfection


The 4D-Nucleofector® 96-well Unit offers a medium-throughput solution suited for screens in 96-well format, the convenient optimization of Nucleofection® Conditions or assay development. For scalable throughput, each of the 96 wells 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 or watch our video about easy transfer of transfection conditions.

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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 1x107 to 1x109 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 allow you to quickly translate research results into a GMP compliant cell therapy manufacturing process. Learn more by visiting our 4D-Nucleofector® LV Unit product page.

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384-well Nucleofector® System: High-throughput Transfection in 384-well Format 


The 384-well 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, including CRISPR Screens. The use of the same conductive polymer in the 384-well Nucleocuvette® plates facilitates transferability of protocols between the 384-well Nucleofector® System and the 4D-Nucleofector® System. Also, the system can be fully integrated into liquid handling systems. Find out more by visiting the 384-well Nucleofector® System product page or watch this video to see the 384-well HT Nucleofector® System in an automated setup.

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Are you interested to learn more about our Nucleofector® Systems? Request further information and one of our specialists will contact you shortly to schedule your personal meeting.
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Bioscience Knowledge Center

Browse our Cell and Transfection, Citation, FAQ and Culture Media Technical Databases.
 
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References:

Maasho et al. (2004) Efficient gene transfer into the human natural killer cell line, NKL, using the Amaxa nucleofection system Journal Immunol Methods, 284(1-2):133-40
Marques and Williams (2005) Activation of the mammalian immune system by siRNAs Nat Biotechnol, 23(11):1399-405
Seki and Rutz (2018) Optimized RNP transfection for highly efficient CRISPR/Cas9-mediated gene knockout in primary T cells J Exp Med, 215(3):985-997
Roth et al. (2020) Pooled Knockin Targeting for Genome Engineering of Cellular Immunotherapies Cell, 181(3):728-744.e21
Kaji et al. (2009) Virus-free induction of pluripotency and subsequent excision of reprogramming factors Nature, 458(7239):771-5
Byrne and Chruch (2015) Crispr-mediated Gene Targeting of Human Induced Pluripotent Stem Cells Curr Protoc Stem Cell Biol, 35(Suppl 35):5A.8.1-5A.8.22
Monjezi et al. (2017) Enhanced CAR T-cell engineering using non-viral Sleeping Beauty transposition from minicircle vectors Leukemia, 31(1):186-194
Georgiadis et al. (2021) Base-edited CAR T cells for combinational therapy against T cell malignancies Leukemia, 35(12):3466-3481