(366e) Translating DNA Origami Nanotechnology to Middle School, High School, and Undergraduate Laboratories

DNA nanotechnology is an emerging field enabling researchers to create nanoscale constructs and tools that have potential to advance many fields. Specifically, DNA origami is a rapidly evolving nanotechnology that allows researchers to create geometrically precise and complex nanostructures for applications in fields such as molecular sensing, robotics, and nanomedicine. The interdisciplinary nature of the development and applications of DNA origami can provide many benefits to students exposed to basic knowledge and methods of the nanotechnology. However, the cost of materials and equipment and complexity of design and characterization of nanostructures limits DNA origami experiments mainly to research focused institutions in graduate level laboratories with prior expertise and well-equipped laboratories. This work seeks to overcome crucial barriers to translating DNA origami methods to educational and non-graduate laboratories such as middle-school and high school science classes and primarily undergraduate institutions.

The most significant barriers are related to resources and time required for DNA origami fabrication. The equipment needed ranges from possibly available (e.g. gel electrophoresis), to unlikely available (e.g. thermocyclers), or impractical (e.g. electron microscopy) for ready access in educational settings. The barrier of time comes from all stages of the process including: 1) Design – which may take up to weeks for novice users to learn the design process or even learn the design software; 2) Fabrication – DNA origami fabrication may take several hours to prepare and up to days for thermal annealing; 3) Analysis – analysis methods like gel electrophoresis typically requires 2-4 hours to setup and run and microscopy imaging is likely impractical for educational settings. In addition to these barriers the complexity of the design and fabrication process are challenging to educational translation. Here, we present a streamlined protocol for fabrication and analysis of static and dynamic DNA origami nanostructures that can be conducted within a 2-hour laboratory course using low-cost equipment that contains readily available materials in educational laboratories and science classrooms.

The first educational experiment module focuses on a static DNA origami nanorod structure that was previously developed for drug delivery applications. The second educational experiment module focuses on a nanomechanical DNA origami structure initially designed to provide a framework and model for deformable, or compliant, nanostructures. These modules provide a protocol for students to analyze structures via gel electrophoresis using classroom-ready gel equipment. The modules include learning tools related to DNA nanotechnology such as charge screening, mechanical deformations, conformational dynamics and free energy landscapes, nanoscale stimulus response, polymerization, and algorithmic design and assembly. Additionally, the nanomechanical DNA origami educational modules provide a framework to illustrate how simple beam bending mechanical models can be used to predict nanostructure properties, as a class extension. The modules provide topics that relate to a variety of subjects including engineering, chemistry, physics, biology, materials science, medicine, and computer science, and hence could be worked into a range of curricula.

Over the long-term, introducing a broad range of students to DNA origami would also have the potential to advance the field due to increased interest and involvement by young students, who may then pursue education, research, or career paths related to DNA nanotechnology. This work focuses on eliminating major barriers that make DNA origami fabrication and experiments challenging to perform in educational laboratories and classroom environments. Here we overcome these barriers to enable the hands-on introduction and use of DNA origami technology in instructional labs and classrooms. Furthermore, introducing students to DNA nanotechnology and related fields can also have the potential to increase interest and future involvement by young students.