A Unique Mammalian Cell Transfection Protocol


At CellEDIT, we deliver for you, CRISPR-Cas complexes directly to where they are needed: 
the nucleus.

Current Challenges in Mammalian Cell Transfection

The successful transfection of cells poses significant challenges, stemming from the intricate nature of the process and the multiple steps involved in introducing external nucleic acids, peptides, or proteins into eukaryotic cells. [1-3] Particularly when dealing with mammalian cell transfection methods, two crucial hurdles must be surmounted.

Amidst the array of existing cell transfection techniques, the single-cell transfection approach utilizing the FluidFM technology stands out for its exceptional merits. This method not only presents a comprehensive solution to circumvent substantial limitations but also offers a set of distinctive advantages for each step of the transfection process. 

Do you need more information about cell transfection methods?

Step 1: Pass the Cell Membrane & Avoid Random Transport, Cytosolic Degradation 

The revolutionary single-cell transfection method offered by the CellEDIT workflow introduces a groundbreaking capability to deliver precise and highly controlled quantities of CRISPR Cas9 Ribonucleoproteins directly into the nucleus. By confining the DNA cutting process exclusively within the nucleus, this technique effectively sidesteps the potential activation of immune responses and cytosolic degradation. As a result, the efficiency of this method surpasses that of conventional approaches, maximizing the desired outcomes with unparalleled precision.

Mammalian Cell Transfection Protocol by CellEDIT_Cytosurge

Step 2: Pass the Nuclear Envelope & Deliver

Distinguished from conventional cell transfection methods, the CellEDIT workflow employs a remarkable transfection technique rooted in the innovative FluidFM technology. This cutting-edge approach accomplishes the delivery of Ribonucleoproteins (RNPs) directly into the nucleus through precise nano-injection, effectively eliminating an entire step from the experimental process. By adopting this bottom-up strategy, the number of procedural steps involved in cell transfection is significantly reduced. Furthermore, since the injection is conducted on individual isolated cells, the need for laborious cell sorting procedures is entirely eliminated. This streamlined methodology streamlines the workflow and enhances the overall efficiency of the transfection process.

Mammalian Cell Transfection Protocol by CellEDIT_Cytosurge

Step 3: Perform Step 1 & 2 while keeping the cell alive

Unlock the potential of your research endeavors with the groundbreaking cell transfection approach offered by the CellEDIT Service. This innovative methodology prioritizes cell viability by employing a non-destructive and meticulously controlled technique to deliver CRISPR complexes directly into the nucleus. Whether your objective is gene knock-in, gene knock-out, or multiplexing, this cutting-edge workflow caters to a diverse array of applications. With CellEDIT Service, you can confidently embark on your scientific journey, knowing that you have access to a revolutionary tool that enhances precision and expands the possibilities of genetic manipulation.

Mammalian Cell Transfection Protocol by CellEDIT_Cytosurge

Cell Transfection As A Service

Resources

On-Demand CellEDIT's CRISPR Cell Line Engineering Webinar

CellEDIT's Service Brochure

CellEDIT's Application Note n*1 - Introducing the CellEDIT Workflow

CellEDIT's Application Note n*2 - Overcoming the hard-to-transfect cell line hurdle 

Open Access Publication featuring CellEDIT:  Antony, Justin S., Anabel Migenda Herranz, Tahereh Mohammadian Gol, Susanne Mailand, Paul Monnier, Jennifer Rottenberger, Alicia Roig‐Merino et al. " Accelerated generation of gene-engineered monoclonal CHO cell lines using FluidFM nanoinjection and CRISPR/Cas9Biotechnology Journal 19, no. 4 (2024): 2300505.

Case Study - Streamlined U2OS Cell Line Modification with the CellEDIT Service Workflow, featuring our CellEDIT Customer: Dr. Kanstantsin Siniuk.

References

[1] Chong, Zhi Xiong, Swee Keong Yeap, and Wan Yong Ho. "Transfection types, methods and strategies: A technical review." PeerJ 9 (2021): e11165.

[2] Kim & Eberwine (2010) Kim TK, Eberwine JH. Mammalian cell transfection: the present and the future. Analytical and Bioanalytical Chemistry. 2010;397(8):3173–3178. doi: 10.1007/s00216-010-3821-6.

[3] Fus-Kujawa, Agnieszka, et al. "An overview of methods and tools for transfection of eukaryotic cells in vitro." Frontiers in Bioengineering and Biotechnology (2021): 634.