Engineering a DNA-based nanodevice to enable cytosolic transport of enzymes

University Of Maryland, College Park, College Park MD

Investigators

Abstract

Abstract Proposal Number: 1402756 P.I.: Muro, Silvia Institution: University of Maryland College Park Title: Engineering a DNA-based nanodevice to enable cytosolic transport of enzymes This project aims at designing tools to enable safe, specific, and efficient access into the interior of live cells, which is elusive to current technologies. This is achieved by leveraging on the material that cells naturally use to store their genetic information and re-engineering this material into tiny devices capable to interact and be transported into cells. The project encompasses characterization of these devices, evaluation of their interaction with live cells, and investigation of their effects. By providing access into live cells, this technology has a very broad potential: it can be used to study how cells function and also to manipulate their activity, which is important to enable research, to exploit cells as factories and machines, and to design new diagnostic and therapeutic tools. This has relevance to advance fundamental knowledge in a variety of disciplines, such as molecular and cellular biology, biomaterials, bioengineering, nanotechnology, etc., and to impact biotechnological production, biopharmaceutical development, and medical applications. The fundamental concepts, methods, and tools developed are also integrated into educational and training components, which involve high school, undergraduate, and graduate students, hence, contributing to training future scientists, engineers, educators, and other workforce. Products of this project are used for scientific divulgation to the specialized and broader community, as well as interaction with industry, to maximize its fundamental and translational impact at a local, national, and global level. The objective of this project is to design and optimize a DNA-based nanodevice to enable safe, specific, and efficient access of biomolecules to the cell interior, which currently remains elusive. This is achieved by exploiting newly discovered properties of DNA, which arise from engineering this natural polymer into architectures different from those available in nature. Briefly, DNA is engineered to acquire a dendrimeric conformation functionalized to carry biomolecules and to target specific cell-surface markers, which renders precise interaction with selected cells, uptake via precise endocytic pathways, and cytosolic delivery with transport to intracellular sites of choice. The three aims explored include characterization of DNA systems with different biophysical properties, evaluation of their interaction with cells, and investigation of their effects in cells. This enables intracellular access of probes, modulatory agents, diagnostics, therapeutics, etc., which has applications in fundamental research, biotechnological production, biopharmaceutical development, and medical applications. By leveraging on diverse disciplines such as cellular biology, biotechnology, bioengineering, nanotechnology, and biopharmaceuticals, this project also contributes tools, methods, and knowledge pertaining to these areas. This is further integrated into educational and training components, which involve high school, undergraduate, and graduate students, contributing to training the future workforce. This award by the Biotechnology, Biochemical, and Biomass Engineering Program of the CBET Division is co-funded by the Instrument Development for Biological Research Program of the Division of Biological Infrastructure.