Today, we introduce you to Kanstantsin Siniuk, whose work focuses on mitochondrial metabolism as a target for developing treatments for mitochondrial and neurodegenerative diseases. Recently, Kanstantsin had the pleasure of collaborating with Cytosurge on a critical project involving the U2OS cell line. This collaboration was essential in validating his findings from two independent sources, contributing to the (hopefully) successful, revision of a paper currently under review.

Kanstantsin Siniuk, PhD, Postdoc in the project group of Dr. Helmut Pospiech at the Leibniz Institute on Aging – Fritz Lipmann Institute (FLI), Jena, Germany.

Our journey with Cytosurge began when I was exploring options for obtaining a modification, a knockout (KO) to be precise, in a specific cell line. We had recently submitted our work to be published in a renowned journal and were asked by the reviewers to obtain additional data to confirm our findings in an independent model system. What ultimately led us to choose Cytosurge was how they efficiently managed the project by getting a CRISPR expert involved so no time was lost. With our project already under revision in a journal, time was of the essence. Cytosurge's timely service was exactly what we needed.  

One of the main reasons we opted for Cytosurge was their flexibility in working with, U2OS cell line that was used for the previous experiments. Unlike other companies, which often require the purchase of a standard cell line, Cytosurge allowed us to send our cell line for modifications. This was crucial because it ensured that the modified cells would behave consistently with our existing data. Moreover, Cytosurge had prior experience engineering such cell lines. Logistics significantly influenced our decision, as Cytosurge's European location offered a logistical advantage that streamlined and accelerated the entire project. This facilitated easier coordination, reduced shipping times and costs, and minimized customs bureaucracy.

Communication was another key factor that led to the project's success and prompt execution as mentioned before. Working with Dr. Tobias Beyer, CSO and Application Scientist, and Dr. Gorka Santos, CellEDIT Sales Manager, at Cytosurge, was a positive experience. Their understanding of scientific terminology and quick responses saved us time and helped us avoid potential errors. This level of expertise and timely communication is something we found lacking in other companies we contacted for this project, where interactions were often limited to sales managers who lacked in-depth scientific knowledge and replies were received after many days or even weeks. 

The design of the edits was tricky due to overlapping regions with other genetic elements, including coding and noncoding exons of another gene, as well as an antisense transcript. Thus, the need for precise KO of more than 2 bp – or complete deletion of the binding site – to ensure that the regulation of the gene by p53 was completely abolished. However, Cytosurge managed to achieve this with accuracy.

The results were as expected; the KO led to the gene no longer being upregulated by p53 induction, confirming the direct impact of the binding site.

Overall, the collaboration with Cytosurge was smooth and efficient. Their scientific knowledge, quick responses, flexibility, and competitive pricing made them an excellent partner for our project. The quality of service and the ability to send and receive modified cell lines quickly are definite advantages.

In conclusion, I would gladly hire Cytosurge for future projects. Their expertise, communication, and ability to deliver timely results make them a valuable partner in the field of genetic research. Their single-cell technology and intranuclear injection method are a beneficial approach for future projects that might involve more complex modifications. As long as their pricing remains competitive, I foresee a continued collaboration with Cytosurge in our future endeavors.