Boost your transfection success with these proven expert tips

insights from industryHinnah CampwalaPrincipal ScientistSartorius

In this interview, News Med talks to Hinnah Campwala about the pivotal role of transfection in modern research and drug discovery.

Transfection is a tool that sees widespread use across a diverse range of research and drug discovery areas. Transfection facilitates the controlled introduction of DNA or RNA, creating new avenues in the exploration of gene functions and cellular pathways without the need to use viral vectors.

Image Credit: Billion Photos/Shutterstock.com

This interview with Hinnah Campwala, Principal Scientist in Sartorius’s Cell Biology Development Group, looks at some of the many advantages and best practice approaches to transfection.

What are the differences between DNA and mRNA transfection?

The choice between DNA and mRNA is an important consideration in transfection, as each offers distinct advantages.

DNA transfection is generally preferred for long-term, stable expression and for integration into the host genome—useful when generating stable cell lines. It’s also beneficial for CRISPR editing and applications that require co-expression of multiple genes.

mRNA transfection, by contrast, enables faster protein expression and avoids the need for nuclear entry and transcription. This makes it well suited for time-sensitive studies like cell-reporter assays, particularly with difficult-to-transfect cells such as primary immune cells.

DNA is typically more stable and cost-effective than mRNA, making it a common choice across many studies. However, mRNA transfection is especially effective when rapid protein expression is needed, especially in cell types that are harder to transfect.

What are some of the most common challenges associated with hard-to-transfect cells?

Working with hard-to-transfect cells like mesenchymal stem cells or T cells is often one of the most challenging aspects of transfection. This is because these cell types are typically less receptive to standard transfection protocols, resulting in high cytotoxicity, low efficiency, and inconsistent results.

I have found that selecting the right reagent is key, with specialized reagents able to provide targeted delivery while helping to reduce cytotoxic effects.

For example, I have had success working with Polyplus reagents to transfect human mesenchymal stem cells and T cells, for example, jetOPTIMUS® for DNA and jetMESSENGER® for mRNA.

The scientific team from Polyplus has also been extremely supportive in troubleshooting these complex projects.

What are some of the most recent advances in chemical transfection?

Chemical transfection methods have seen significant advances in recent years, with improvements in formulations that boost both cell viability and transfection efficiency coming to the fore. These innovations are particularly valuable in high-throughput applications or research working with sensitive cell types.

For example, lipid-based reagents now boast transfection efficiencies of more than 80 % with high cell viability, making them an ideal option in a diverse array of settings.

Lipid-based reagents such as liposomes and lipid nanoparticles can now offer targeted delivery alongside enhanced uptake in hard-to-transfect cells. New formulations improve target specificity while minimizing off-target effects, including cell-penetrating peptides and ligand-conjugated reagents.

What are some of the most important tips when working with transfection?

Maximizing transfection efficiency while minimizing cell stress is key, and this requires attention to detail and an understanding of the transfection process.

For example, it is important to select a reagent that is compatible with your nucleic acid and cell type, adjusting variables like incubation time, concentration, and temperature as required.

It is also advisable to use actively dividing, contamination-free cells that have been seeded at the recommended density. Properly stored high-quality nucleic acids are key, and it is important to avoid freeze-thaw cycles in order to maintain nucleic acid integrity.

Cell health and transfection efficiency should be tracked, with media replaced or refreshed as needed in order to help minimize cytotoxicity. Where possible, cells should be monitored in real time, leveraging high-throughput tools for live-cell analysis and screening cytometry to track efficiency and adjust conditions as required.

What are some of the most common application fields where Polyplus’ transfection technology is used?

Now part of Sartorius, Polyplus Transfection’s range of innovative and high-quality nucleic acid solutions has been specifically tailored for use in research, bioproduction, and gene therapy applications.

Researchers undertaking gene function studies can leverage transfection to manipulate gene expression, allowing them to better understand cellular processes and functions.

By tracking specific genes as part of reporter-gene studies, scientists can achieve real-time monitoring of cell behavior, gene expression, and protein interactions.

Transfection is also instrumental in validating gene targets and screening therapeutic compounds as part of therapeutic discovery.

How do you see transfection technology evolving in the future?

Transfection technology is set to become even more precise and scalable, as next-generation reagents such as advanced lipid nanoparticles and biodegradable polymers contribute to enhanced transfection precision and safety.

These advances will also make transfection more versatile and applicable to different fields, improving gene-editing accuracy and opening doors to new therapeutic applications.

These innovations are also expected to work alongside tools like CRISPR-Cas9, prime editing, and base editing, with transfection playing an increasingly pivotal role in therapeutic development and personalized medicine.

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Acknowledgments

Produced from materials originally authored by Sartorius Corporation.

About Hinnah Campwala

Hinnah Campwala is a Principal Scientist in the (Product Development) Reagent and Consumables Group at Sartorius. With a background in drug discovery and immune cell biology, she specializes in the development of innovative reagents and applications to support researchers working with Sartorius’s advanced bioanalytical tools.

About Sartorius

Sartorius is a leading international pharmaceutical and laboratory equipment supplier. With our innovative products and services, we are helping our customers across the entire globe to implement their complex and quality-critical biomanufacturing and laboratory processes reliably and economically.

The Group companies are united under the roof of Sartorius AG, which is listed on the Frankfurt Stock Exchange and holds the majority stake in Sartorius Stedim Biotech S.A. Quoted on the Paris Stock Exchange, this subgroup is comprised mainly of the Bioprocess Solutions Division.

Innovative technologies enable medical progress

A growing number of medications are biopharmaceuticals. These are produced using living cells in complex, lengthy and expensive procedures. The Bioprocess Solutions Division provides the essential products and technologies to accomplish this.

In fact, Sartorius has been pioneering and setting the standards for single-use products that are currently used throughout all biopharmaceutical manufacturing processes.

Making lab life easier

Lab work is complex and demanding: Despite repetitive analytical routines, lab staff must perform each step in a highly concentrated and careful way for accurate results.

The Lab Products and Services Division helps lab personnel excel because its products, such as laboratory balances, pipettes and lab consumables, minimize human error, simplify workflows and reduce physical workloads.


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