One of the key elements of the MITGEST Doctoral Network is the mobility of researchers, allowing our doctoral candidates (DCs) to transfer between different institutions, including academia and industry. So-called secondments have already started within the network and we are proud to announce the successful start of a research secondment of DC3, Dory Vergallo, who transferred from the host institution – Radboud Center for Mitochondrial Medicine (Nijmegen, Netherlands) to baseclick GmbH (Neuried, Germany). This exceptional opportunity allows for collaboration with esteemed professionals in the field of Click Chemistry technology, specifically Dr. Thomas Frischmuth and Dr. Fabio Spada. In the following article, readers will gain insights directly from Dory (DC3), who will share a personal account of this enriching experience.

What is Ribonucleic acid and why it is important?

Ribonucleic acid (RNA) is a molecule that is essential for most biological functions, either by performing the function itself or by forming a template for the production of proteins. Mitochondrial RNA plays a crucial role in cellular function, particularly in energy production and metabolism. By developing efficient methods to tag nascent mitochondrial RNA, we open doors to a better understanding of its dynamics and regulation, which is fundamental for unravelling the mysteries of mitochondrial biology and its implications in various diseases.

About my research on mitochondrial RNA

During my time at Basecklick, one of the key partners of the MITGEST project, I worked on making it easier to label and study newly made mitochondrial RNA. Baseclisk has developed an effective method called Click Chemistry to tag DNA and RNA. I brought RNA samples that contained a special molecule called 5-Ethynyl Uridine (5-EU). This molecule can be added to cell cultures and gets incorporated into newly made RNA. To detect and label 5-EU, we use a technique called Click-chemistry, specifically the Copper-catalysed Azide-Alkyne Cycloaddition (CuAAC) reaction. This reaction helps 5-EU bind to another molecule called azide biotin. We can then use magnetic beads to capture the biotin-tagged RNA.

What is the challenge and how do we address it?

However, Click-chemistry needs copper, which can be harmful in large amounts. Imagine copper as a helper that makes the reaction happen, but too much of it can create harmful by-products that damage RNA. This damage makes it harder to get good-quality RNA for our studies.

To solve this, we tried using a different type of biotin linked to picolyl azide, which needs less copper. This reduced the damage to RNA, but we still saw some degradation. There are also copper-free methods (called SP Click chemistry), but they only work outside living cells and aren’t useful for our purposes.

Our main goal is to improve the Click-chemistry for 5-EU tagged RNA by using as little copper as possible, to get better quality RNA. Baseclick is testing new reagents called azide plus, which form strong bonds with copper and make the reaction happen faster and with much less copper.

My work involves testing these new molecules with 5-EU labelled mitochondrial RNA and comparing the results with those from picolyl-biotin-azide. I aim to develop a better method for isolating newly made RNA, which will help us study how genes are regulated in cells.


Taking out from the experience

Reflecting on this experience, I’m overflowing with gratitude for the support and mentorship I’ve already received during my short time here. The collaborative atmosphere at baseclick has truly fueled creativity and innovation, pushing me to push the boundaries of my research and make meaningful contributions to the field.

In closing, I urge fellow researchers and enthusiasts to embrace opportunities for collaboration and knowledge exchange. Together, we can unravel the complexities of mitochondrial biology and pave the way for advancements in biomedical research. As I embark on the next phase of my doctoral journey, I am excited to apply the insights gained from my time at baseclick and continue exploring the fascinating realm of mitochondrial RNA biology.

The news item was written by MITGEST DC3 Dory Vergallo.

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