Principal Investigator Carnegie Mellon University, United States
Introduction:: Our work focuses on developing a novel RNA therapeutic nanoparticle formulation that thermally stabilizes RNA, protecting it from degradation and potency loss during the supply chain. This nanoparticle technology utilizes Metal-Organic Frameworks (MOFs) to encapsulate, stabilize, and deliver RNA, as shown with DNA and proteins [1]. A thermally stable RNA therapeutic removes the need for cold chain transport allowing rural and low-resource locations easier accessibility to life-saving medicine.
Materials and Methods:: RNA encapsulating nanoparticles are synthesized using non-aqueous solutions of Zinc Nitrate and 2-Methyl Imidazole. Particles are then prepared and stored in various thermal environments. Particles are then resuspended. HEK 293t Cells are transfected with the RNA encapsulating particles and imaged. Fluorescent measurements are obtained using a plate reader. Particles are characterized using scanning electron microscopy, zeta sizer, and powder x-ray diffraction. Agarose gels were used to confirm RNA loading efficiency and RNA stability.
Results, Conclusions, and Discussions:: Metal-organic framework(MOF)-RNA particles are about 60 - 200 nm in size, and RNA encapsulation causes no structural changes to the encapsulating MOF material. This MOF system has an RNA loading efficiency of 91.5%. The encapsulated RNA was stable inside the MOF nanoparticles for two months, with some loss in transfection efficiency compared to controls stored at -80˚C and 4˚C. Overall, our novel RNA encapsulation method is promising as we take steps to understand how to improve RNA stabilization at room temperature.
Acknowledgements (Optional): : National Science Foundation GFP
References (Optional): :
[1] Liang, K., Ricco, R., Doherty, C. M., Styles, M. J., Bell, S., Kirby, N., Mudie, S., Haylock, D., Hill, A. J., Doonan, C. J., & Falcaro, P. (2015). Biomimetic mineralization of metal-organic frameworks as protective coatings for biomacromolecules. Nature Communications, 6(1), 7240. https://doi.org/10.1038/ncomms8240
