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Celebrating 20 Years of Nucleofector® Technology

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2021 marks the 20th anniversary of the Nucleofector® Technology, a non-viral cell transfection method which has been cited more than 10,000 times. In the 20 years since its development, the technology has provided researchers with a tool to efficiently modify hard-to-transfect cells, fuelling research in fields ranging from drug discovery to cell and gene therapy.

To celebrate this milestone, Technology Networks spoke with Archie Cullen, senior vice president and head of bioscience solutions, Lonza. In this interview, Archie discusses the evolution of the Nucleofector® Technology, highlighting how it has supported and propelled basic and applied research. He also explores some of the most recent developments in the Nucleofector® Platform’s exciting journey.

Anna MacDonald (AM): Can you tell us about the history of Nucleofector® Technology?

Archie Cullen (AC):
Nucleofector® Technology was invented more than 20 years ago in response to the challenges scientists were facing from hard-to-transfect primary cells and cell lines. Traditional transfection methods, such as lipofection or electroporation, were failing to deliver acceptable transfection efficiency and cell viability, especially for immune cells. Nucleofector® Technology was specifically created to solve these problems.

Over its 20-year history, the platform has evolved through continuous development to further improve results and service new demands from researchers all over the world. The technology has evolved to support new applications, from CRISPR screening to cell and gene therapy. With the introduction of consumables fabricated from conductive polymers, the Nucleofection® Procedure creates even gentler conditions, preventing the release of Al3+ ions during transfection that can cause cell death post-transfection. These new Nucleocuvette® Consumables paved the way for the 4D-Nucleofector® Platform, designed to support easy scale-up and transfer of transfection conditions into any given format.

Lonza has a long history of fast-paced and continuous innovation, listening to its customers and adjusting technology to better meet the industry’s changing needs. The Nucleofector® LV Unit is Lonza’s latest module designed for use with good manufacturing practice (GMP) consumables, to achieve transfection of high cell numbers for cell and gene therapy.

AM: How does the Nucleofector® Technology compare to other methods of transfection?

Nucleofector® Technology is an advanced form of electroporation, combining the Nucleofector® Device, specialized Nucleofector® Solutions, and optimized protocols to create a unique transfection system. Compared to other transfection methods, such as lipofection or chemical transfection, results show better reproducibility, performance, transfection efficiency and higher cell viability for hard-to-transfect cells.

Standard electroporation techniques create a short-lived electric field to make pores in the cell membrane, allowing DNA or other substrates to be delivered into the cytoplasm. Conversely, the Nucleofection® Procedure delivers DNA directly into the nucleus. Researchers can, therefore, accelerate research without waiting for the nucleus to break down during cell division and uptake the introduced DNA. Gene expression can be observed as soon as three hours after transfection, and transfection efficiencies can exceed 90%.

AM: In what ways has Nucleofector® Technology helped create novel opportunities for basic and applied research?

The versatility of the Nucleofector® Platform enables scientists to scale their research depending on their needs. The technology supports a broad range of transfection applications, from basic research with low to medium throughput and cell numbers to largescale applications in 96-well or 384-well formats, such as CRISPR library screens. The large-volume format enables transfection of up to two billion cells at once, making it ideally suited for autologous non-viral cell and gene therapy. The substrate-independent transfection conditions support complex experimental setups with multiple knockouts at once, as well as transfection of ribonucleoproteins (RNPs) and peptides. Furthermore, with Lonza’s comprehensive cell biology portfolio, customers are supported with the primary cells and cell culture media that complement Nucleofector® Technology. Integration with the Cocoon® Platform helps support the next generation of patient-specific cell therapy manufacturing.

AM: How has Nucleofector® Technology evolved over the past 20 years to meet the changing needs of scientists?

The Nucleofector® System has already supported a plethora of innovations and has been used for more than 10,000 peer-reviewed publications since its launch. The newly launched 4D-Nucleofector® Platform offers further flexibility, ease-of-use and safer transfection conditions. The modular concept facilitates versatile setup tailored to the specific application needs of researchers. GMP consumables and the conductive polymer allow for safe, non-viral transfection for cell and gene therapy applications. The 96-well module and 384-well HT-Nucleofector® System are routinely used for CRISPR screening on a primary cell background. The easy-to-use kit provides optimal transfection conditions and transferability so that once established, transfection conditions can be directly used in literally any other format, providing researchers with a toolbox fully equipped for their application needs.

AM: Can you discuss the next stage of development for Nucleofector® Technology? What improvements can users expect?

The new generation of the 4D-Nucleofector® Platform has recently launched. Customers can now benefit from an improved user experience and full integration of the 96-well unit for medium-throughput applications. This new release comes complete with a new design and user-friendly software for experiment setup and data management. 4D-Nucleofector® Platform customers can expect continuing high standards of performance supported with technology that is even easier to use.

Archie Cullen was speaking to Anna MacDonald, Science Writer for Technology Networks.