Revolutionising RNA Chip Production: New Building Blocks Increase Efficiency and Speed

An international research team has developed a new version of RNA building blocks, significantly enhancing the production speed and efficiency of RNA chips.

The University of Vienna, in collaboration with the Max Mousseron Institute for Biomolecules at the University of Montpellier, has made a significant breakthrough in RNA technology. The new RNA building blocks exhibit higher chemical reactivity and photosensitivity, drastically reducing production times and increasing efficiency. Published in Science Advances, this development has far-reaching implications for biotechnology and medical research.

Overcoming Stability Challenges in RNA Microarrays

RNA, unlike DNA, presents unique challenges due to its lower stability, making it difficult to adapt DNA microarray technology for RNA. Despite the significant role of RNA in biological processes and its potential in medical applications, the synthesis of stable RNA microarrays has been problematic. Traditional methods have faced issues with long production times, low yields, and poor stability.

The method for chemical synthesis of RNA, developed 40 years ago, uses phosphoramidite chemistry, allowing any sequence to be assembled from RNA building blocks. This method is crucial in the production of microarrays, where millions of unique sequences can be analysed simultaneously on a solid surface. The University of Vienna first demonstrated high-density RNA chip production using photolithography in 2018. This technique uses light to precisely position building blocks on a surface through a photochemical reaction. However, the method’s initial version was limited by inefficiencies and stability issues.

The research team has now introduced a new generation of RNA building blocks, which significantly improve the efficiency and speed of RNA chip synthesis. The novel building blocks are equipped with propionyloxymethyl (PrOM) protecting groups, enhancing their chemical reactivity and photosensitivity. This innovation has made the synthesis process twice as fast and seven times more efficient.

According to Jory Lietard, Assistant Professor at the Institute of Inorganic Chemistry, “Making RNA microarrays containing functional RNA molecules was simply out of reach with our earlier setup, but it is now a reality with this improved process using the PrOM protecting group.”

Applications and Future Prospects

The newly developed RNA chips have demonstrated their potential in screening millions of candidate RNA sequences. The publication details a study of RNA aptamers, small oligonucleotides that specifically bind to target molecules. The researchers synthesised thousands of variants of two “light-up” aptamers, which produce fluorescence upon binding to a dye. This approach allows for the simultaneous acquisition of data on all variants, paving the way for identifying improved aptamers with better diagnostic properties.

Tadija Kekić, a PhD candidate in Lietard’s group, highlights the significance of this advancement: “High-quality RNA chips could be especially valuable in the rapidly growing field of non-invasive molecular diagnostics. New and improved RNA aptamers are critically sought after, such as those that can track hormone levels in real-time or monitor other biological markers directly from sweat or saliva.”

The enhanced efficiency and speed of RNA chip production can lead to significant advancements in various fields, including molecular diagnostics, personalised medicine, and biotechnological applications. Researchers and laboratory technicians can now utilise these high-quality RNA chips to conduct more comprehensive and efficient analyses, potentially accelerating the discovery and development of new medical diagnostics and treatments.

For more information, please refer to the full study “Accelerated, high quality photolithographic synthesis of RNA microarrays in situ” published in Science Advances.