Invention Description
In the rapidly evolving field of live-cell analysis, recent innovations have enabled the sensing and quantification of specific intracellular nucleic acid targets, such as messenger RNA (mRNA) and other RNA species, in viable cells without requiring cell lysis or destruction of the sample. This capability is critical for drug discovery and personalized medicine, as it allows researchers to observe complex cellular behaviors in real-time. However, a major commercial hurdle remains: the conventional sensors for detecting these intracellular targets often cause cell stress or toxicity when integrated into the cell membrane for prolonged periods. This extended binding frequently compromises the cell's health, rendering the biological samples unsuitable for essential downstream processes like high-speed Fluorescence-Activated Cell Sorting (FACS). Consequently, researchers are often forced to choose between capturing high-quality data and preserving the life of the cell. This bottleneck limits the viability of cells for critical workflows and hinders the scalability of sequential analysis technologies. Addressing this cytotoxicity is essential to unlocking the full market potential of advanced diagnostic platforms and live-cell research.
Researchers at ASU have developed a method for efficiently removing DNA based transmembrane sensors from cell membranes, that binds with a specific intracellular nucleic acid targets (DNA/RNA) sequence. These transmembrane DNA nanosensors are internally modified with cholesterol for membrane anchoring and fragments are conjugated with fluorescent and quencher labels to allow for signal detection upon binding with the target nucleic acids. Removal of cholesterol-modified sensors would allow for the reduction of cytotoxicity in sensor-based cellular applications. The method uses cyclodextrin derivatives to efficiently remove cholesterol-modified transmembrane sensors by forming a host-guest complex, facilitating their removal from the cell membrane and ensuring clearance of the sensors from the cell membrane.
Potential Applications
• Biomedical research involving live-cell RNA biomarker detection
• Clinical diagnostics with cell sorting technologies (FACS, MACS)
• Development of molecular diagnostic devices and kits
• Pharmaceutical screening and personalized medicine
Benefits and Advantages
• Significantly reduced cytotoxicity compared to existing methods
• Highly effective with over 90% sensor clearance
• Simple and efficient sensor removal process
• Versatile and compatible with live-cell RNA detection techniques
• Enhances cell viability and sorting accuracy