Multifunctional RNA/DNA nanoassemblies for therapeutic activation

The National Institute of Allergy and Infectious Diseases awarded $118,332 to the University of North Carolina at Charlotte on October 9, 2026, under the Allergy and Infectious Diseases Research program (CFDA 93.855) to develop multifunctional RNA/DNA hybrid nanoassemblies capable of simultaneously activating the cGAS-STING and RIG-I immune pathways while incorporating real-time fluorescent monitoring. The approach involves engineering non-functional RNA/DNA hybrids carrying partial agonists that reassociate within cells to release active DNA and RNA duplexes, with hybrid reassociation controlled and monitored using split fluorogenic aptamers that emit fluorescence upon activation. This controlled-release mechanism prevents premature immune activation and enables precise tracking of pathway activation processes. The research bridges existing work in DNA and RNA duplexes for pathway activation and fluorogenic aptamers for real-time tracking into a single platform for conditional immune activation, offering applications for targeted drug delivery and immune modulation. Performance occurs in Charlotte, North Carolina, with a period of performance extending from October 9, 2026, through January 31, 2029.

Abstract

The immune system has evolved complex pathways to protect the body from harmful intruders, utilizing pattern recognition receptors (PRRS) such as cgas-sting and RIG-I to detect foreign pathogens and abnormal cells, triggering essential immune responses. PRRS serve as the body's early detection system, recognizing specific patterns associated with infections or stress. However, the precise and controlled activation of these pathways remains a significant challenge, largely due to the absence of technologies that allow for conditional activation. This proposal aims to address this GAP by developing RNA/DNA hybrid nanoassemblies capable of simultaneously activating the cgas-sting and RIG-I pathways, while incorporating real-time monitoring through a fluorescent readout. This dual functionality will not only enable precise tracking of activation processes but also provide valuable insights for optimizing therapeutic strategies by improving the timing of intervention. The approach involves introducing non-functional RNA/DNA hybrids, each carrying a partial agonist for either cgas or RIG-I, which can reassociate within cells to release active DNA and RNA duplexes. The hybrid reassociation is controlled and monitored using split fluorogenic aptamers, which emit fluorescence upon activation. By utilizing the controlled release mechanism, this strategy prevents premature activation, ensuring immune responses are triggered only when needed. This strategy builds on existing research in DNA and RNA duplexes for pathway activation and fluorogenic aptamers for real-time tracking, integrating these elements into a single platform for conditional immune activation. By harnessing the versatility of RNA/DNA hybrid nanoassemblies, which can be engineered through rational nucleic acid design, this approach offers a promising platform for targeted drug delivery and immune modulation. Overcoming the challenge of simultaneously activating both the cgas-sting and RIG-I pathways will be key to maximizing the potential of nucleic acid-based therapies and improving treatment outcomes. Precise immune activation will be crucial for controlling responses in diseases such as cancer and infections. This technology holds significant potential for therapeutic applications, aiming to enhance the precision of immune pathway activation, minimize off-target effects, and pave the way for more effective and personalized treatments.