By Dave DeFusco
Two 鶹ýӳ researchers have been awarded a $600,000 grant from the National Science Foundation to solve a longstanding biological mystery: how RNA molecules, which play a crucial role in cell function and organism development, are transported and localized within cells.
Dr. Irina Catrina, associate professor of chemistry, and Dr. Josefa Steinhauer, professor of biology, who both teach in Yeshiva College, received the grant, “RUI: Tools4Cells: Identifying Links Between Intracellular Vesicle Transport and Long-range RNA Trafficking During Drosophila melanogaster Oogenesis,” as part of the NSF’s Research in Undergraduate Institutions program. Their research was seed-funded by the Katz School of Science and Health’s Faculty Research Initiative.
“The mislocalization of RNA can have catastrophic effects,” said Dr. Catrina, who is principal investigator of the project, “including developmental disorders and lethality, making this a high-impact area of study.”
Over the last 50 years, scientific tools, such as specific antibodies, immunofluorescent staining and molecular fluorescent tags, have allowed researchers to map the distribution of thousands of proteins in cells; however, understanding of RNA localization and transport remains limited by comparison. Although a handful of RNA distribution patterns have been studied, many RNAs remain uncharacterized. Furthermore, even when RNA distribution is understood, the mechanisms behind their transport within cells are often unknown.
To address the technical challenges of RNA detection, the researchers are developing computational and experimental tools that will allow them to study RNA transport and localization in real time. A key goal is the creation of a Python-based software package that will simplify the design of custom RNA probes, making this technology more accessible to the broader research community.
These probes, used for detecting RNA molecules, are essential for visualizing where RNAs are located in live cells. The software will integrate the researchers’ previous work on probe design and will provide flexible options for targeting RNA from any organism. By using mathematical models that account for variables like target structure, probe chemistry and environmental factors, such as temperature, the software will rank potential probe candidates for optimal efficiency.
Once the RNA probes have been developed, the researchers will use them to co-visualize RNA and proteins in vivo, or inside the living organism. The research will focus on the fruit fly, Drosophila melanogaster, a common model organism used in genetics and cell biology due to its relatively simple biology and well-mapped genome. Specifically, the researchers will investigate maternal mRNAs, or RNA molecules deposited in the egg by the mother, during oogenesis, the process of egg cell formation. These mRNAs play a critical role in the early development of the embryo, ensuring that cells differentiate into the correct tissue types.
RNAs in the fly egg will be co-visualized with vesicles, small subcellular structures well known for their role in transporting proteins. Only recently have vesicles been linked to RNA trafficking, opening new possibilities for understanding how RNAs move within cells.
By illuminating RNA molecules and simultaneously Rab proteins, which are involved in vesicle transport, they aim to show that vesicles help transport maternal mRNAs within the developing egg cell. Intriguingly, Rab proteins, like Rab5, Rab7 and Rab11, have been linked to RNA transport in neurons, but it remains unknown how widespread this mechanism is across other cell types.
“The findings from these studies could have broad implications, shedding light on RNA trafficking mechanisms that may apply to other species, including humans,” said Dr. Steinhauer, the project’s co-principal investigator.
These experiments will provide proof of principle for the software tools and allow for their optimization and validation, while also yielding important insights into how RNA trafficking is accomplished in the fly egg.
“Our research has the potential to significantly advance our understanding of fundamental cellular processes, with wide-ranging applications,” said Dr. Catrina. “All cells rely on the transport and localization of RNA for proper function, meaning the tools and insights generated from this work could be applied across various organisms and cell types. In particular, these findings may help inform future research into diseases where RNA localization goes awry, such as in certain neurodegenerative conditions.”
