An acoustic liquid handling robot for combinatorial assembly of large DNA nanostructures
Abstract
We propose using a Labcyte Echo 525 acoustic liquid handling robot (i.e. the Echo) to lower barriers to assembling larger (i.e. 0.5~1000 gigadaltons) DNA nanostructures of arbitrary design. The Echo will be used to combine thousands of oligonucleotides into huge DNA nanostructures significantly larger than what methods in DNA nanotechnology routinely allow. This equipment supersedes previous generations of liquid handlers since it uses acoustic waves to transfer liquids. As such, the Echo rapidly and precisely transfers nanoliter volumes without using consumables such as pipette tips. We will leverage the Echo liquid handler to assemble a multitude of large DNA nanostructures using two distinct assembly methods we have developed with DoD support. In one approach, the Echo will be used to manufacture DNA origami ~slats~ that assemble into 1D, 2D, and 3D crystals, as well as finite nanostructures with hundreds of components. In the other approach, the Echo will be used to prepare combinations of tens of thousands of DNA ~bricks~ into arbitrary 3D objects. The Echo will allow us to realize the full potential of both approaches since it overcomes two key limitations of existing pipetting tools. Firstly, it will virtually eliminate user time to manually pipette mixtures of DNA strands. Single nanostructures derived by either method require one-full-day to weeks to pipette DNA strands together. By contrast, the Echo automatically performs these operations over minutes to hours with negligible user labor. This will hugely shorten the design-build-test cycle to assemble large nanostructures, allow further refinement of each approach, and provide a means to test many pertinent scientific questions. Secondly, the Echo will substantially lessen reagent and consumable expenditures. The particular unit here transfers volumes as small as 25 nanoliters, which will enable us generate more DNA nanostructures from a finite library of synthetic DNA. Furthermore, the Echo eliminates the need for pipette tips. The cost savings are equal to the price of the Echo with assembly of only 200 nanostructures, when each are composed of tens-of- thousands of DNA strands. Taken together, the Echo will greatly lower barriers to assembling large DNA nanostructures. This will enable us to use such nanostructures for myriad applications including therapeutics, plasmonic metamaterials, and biophysical tools. More broadly, the Echo will also bolster our other DoD supported research in gene synthesis and synthetic biology. APPROVED FOR PUBLIC RELEASE.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- May 23, 2019
- Source ID
- N000141912345
Entities
People
- William M. Shih
Organizations
- Office of Naval Research
- President and Fellows of Harvard College
- United States Navy