title: Homogeneous Entropy-Driven Amplified Detection of Biomolecular Interactions.
abstract: While a range of artificial biochemical circuits is likely to play a significant role in biological engineering, one of the challenges in the field is the design of circuits that can transduce between biomolecule classes (e.g., moving beyond nucleic acid only circuits). Herein, we design a transduction mechanism whereby a protein signal is transduced into an amplified nucleic acid output using DNA nanotechnology. In this system, a protein is recognized by nucleic acid bound recognition elements to form a catalytic complex that drives a hybridization/displacement reaction on a multicomponent nucleic acid substrate, releasing multiple target single-stranded oligonucleotides in an amplified fashion. Amplification power and simple one-pot reaction conditions lead us to apply the scheme in an assay format, achieving homogeneous and rapid (∼10 min) analyte detection that is also robust (operable in whole blood and plasma). In addition, we demonstrate the assay in a microfluidic digital assay format leading to improved quantification and sensitivity approaching single-molecule levels. The present scheme we believe will have a significant impact on a range of applications from fundamental molecular interaction studies to design of artificial circuits in vivo to high-throughput, multiplexed assays for screening or point-of-care diagnostics.
authors.affiliation: Department of Bioengineering, ‡Department of Pathology & Laboratory Medicine, §Department of Electrical Engineering, ∥California NanoSystems Institute, and ⊥Jonsson Comprehensive Cancer Center, University of California, Los Angeles , Los Angeles, California 90095, United States.
authors.forename: Donghyuk
authors.initials: D
authors.lastname: Kim
authors.affiliation: Department of Bioengineering, ‡Department of Pathology & Laboratory Medicine, §Department of Electrical Engineering, ∥California NanoSystems Institute, and ⊥Jonsson Comprehensive Cancer Center, University of California, Los Angeles , Los Angeles, California 90095, United States.
authors.forename: Omai B
authors.initials: OB
authors.lastname: Garner
authors.affiliation: Department of Bioengineering, ‡Department of Pathology & Laboratory Medicine, §Department of Electrical Engineering, ∥California NanoSystems Institute, and ⊥Jonsson Comprehensive Cancer Center, University of California, Los Angeles , Los Angeles, California 90095, United States.
authors.forename: Aydogan
authors.initials: A
authors.lastname: Ozcan
authors.affiliation: Department of Bioengineering, ‡Department of Pathology & Laboratory Medicine, §Department of Electrical Engineering, ∥California NanoSystems Institute, and ⊥Jonsson Comprehensive Cancer Center, University of California, Los Angeles , Los Angeles, California 90095, United States.
authors.forename: Dino
authors.initials: D
authors.lastname: Di Carlo
mesh_terms: D004247:DNA; D019277:Entropy; D036103:Nanotechnology; D009693:Nucleic Acid Hybridization; D009696:Nucleic Acids
chemical_list: D009696:Nucleic Acids; D004247:DNA
keywords: DNA-based molecular machinery; amplified assay; biomolecular interaction; digital assay; homogeneous assay; protein-to-nucleic acid transduction