News | UMass Amherst Develops Three-Color Fluorescence Imaging for Real-Time RNA Visualization in Living Cells
Chemists at the University of Massachusetts Amherst (UMass Amherst) have developed a new three-color fluorescence imaging method that simultaneously tracks multiple messenger RNAs (mRNAs) in living mammalian cells in real time. The technique labels different types of RNA with different fluorescent colors, allowing researchers to observe their dynamic behavior inside cells. It provides a key tool for understanding RNA, a fundamental molecule of life. The findings were recently published in Nature Methods.
RNA plays an essential yet incompletely understood role in life. It not only carries information between DNA and proteins and directs cells to produce proteins, but also regulates whether genes are switched on or off and helps organize and shape cellular structures. Abnormal RNA function is closely associated with many diseases, making it a major challenge in the life sciences to study RNA without disrupting its natural state.
“We are curious about the many functions of RNA,” said corresponding author Jiahui ‘Chris’ Wu, assistant professor of chemistry at UMass Amherst. “The real question is how to study them. The ideal approach is to observe them directly in living cells, but RNA molecules are simply too small.”
In fluorescence microscopy, scientists generally add fluorescent markers to target RNA so it stands out against the complex cellular background. One widely used technique is the “RNA hairpin method,” which fuses a fluorescent protein to a specific RNA for imaging. However, the fluorescent signal is often continuously active, creating background noise that reduces imaging accuracy.
The UMass Amherst team made a key improvement to this technique. First author Daisy Pham, a graduate student in chemistry, explained that the team designed new fluorescent proteins that light up only after binding to a specific structure in an RNA molecule, substantially reducing background interference. More importantly, the researchers engineered three mutually orthogonal fluorescent proteins emitting green, red and far-red light, allowing RNAs with different functions to be labeled and distinguished simultaneously.
“We can now observe how different types of RNA work at the same time in living cells,” Pham said. “This brings us closer to understanding exactly how each performs its function.”
The team emphasized that the imaging method has been made available to the research community and could become an important new tool for studying RNA function. It enables scientists to obtain more accurate, dynamic information from inside cells without substantially disturbing the RNA itself. The advance expands the capabilities of RNA imaging and provides a methodological foundation for understanding the mechanisms of life and RNA-related diseases.
The study was funded by the U.S. National Institutes of Health (NIH) and the UMass Amherst Institute for Applied Life Sciences (IALS). IALS brings together interdisciplinary expertise from 29 university departments to translate basic research into innovative applications that improve human health and well-being.
News | UMass Amherst Develops Three-Color Fluorescence Imaging for Real-Time RNA Visualization in Living Cells
News | UMass Amherst Develops Three-Color Fluorescence Imaging for Real-Time RNA Visualization in Living Cells
Chemists at the University of Massachusetts Amherst (UMass Amherst) have developed a new three-color fluorescence imaging method that simultaneously tracks multiple messenger RNAs (mRNAs) in living mammalian cells in real time. The technique labels different types of RNA with different fluorescent colors, allowing researchers to observe their dynamic behavior inside cells. It provides a key tool for understanding RNA, a fundamental molecule of life. The findings were recently published in Nature Methods.
RNA plays an essential yet incompletely understood role in life. It not only carries information between DNA and proteins and directs cells to produce proteins, but also regulates whether genes are switched on or off and helps organize and shape cellular structures. Abnormal RNA function is closely associated with many diseases, making it a major challenge in the life sciences to study RNA without disrupting its natural state.
“We are curious about the many functions of RNA,” said corresponding author Jiahui ‘Chris’ Wu, assistant professor of chemistry at UMass Amherst. “The real question is how to study them. The ideal approach is to observe them directly in living cells, but RNA molecules are simply too small.”
In fluorescence microscopy, scientists generally add fluorescent markers to target RNA so it stands out against the complex cellular background. One widely used technique is the “RNA hairpin method,” which fuses a fluorescent protein to a specific RNA for imaging. However, the fluorescent signal is often continuously active, creating background noise that reduces imaging accuracy.
The UMass Amherst team made a key improvement to this technique. First author Daisy Pham, a graduate student in chemistry, explained that the team designed new fluorescent proteins that light up only after binding to a specific structure in an RNA molecule, substantially reducing background interference. More importantly, the researchers engineered three mutually orthogonal fluorescent proteins emitting green, red and far-red light, allowing RNAs with different functions to be labeled and distinguished simultaneously.
“We can now observe how different types of RNA work at the same time in living cells,” Pham said. “This brings us closer to understanding exactly how each performs its function.”
The team emphasized that the imaging method has been made available to the research community and could become an important new tool for studying RNA function. It enables scientists to obtain more accurate, dynamic information from inside cells without substantially disturbing the RNA itself. The advance expands the capabilities of RNA imaging and provides a methodological foundation for understanding the mechanisms of life and RNA-related diseases.
The study was funded by the U.S. National Institutes of Health (NIH) and the UMass Amherst Institute for Applied Life Sciences (IALS). IALS brings together interdisciplinary expertise from 29 university departments to translate basic research into innovative applications that improve human health and well-being.
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