Autonomous dynamic control of DNA nanostructure self-assembly

Biological cells routinely reconfigure their shape using dynamic signalling and regulatory networks that direct self-assembly processes in time and space, through molecular components that sense, process and transmit information from the environment. A similar strategy could be used to enable life-like behaviours in synthetic materials. Nucleic acid nanotechnology offers a promising route towards this goal through a variety of sensors, logic and dynamic components and self-assembling structures. Here, by harnessing both dynamic and structural DNA nanotechnology, we demonstrate dynamic control of the self-assembly of DNA nanotubes—a well-known class of programmable DNA nanostructures. Nanotube assembly and disassembly is controlled with minimal synthetic gene systems, including an autonomous molecular oscillator. We use a coarse-grained computational model to capture nanotube length distribution dynamics in response to inputs from nucleic acid circuits. We hope that these results may find use for the development of responsive nucleic acid materials, with potential applications in biomaterials science, nanofabrication and drug delivery.Nucleic acid nanotechnology offers a promising route towards the design and synthesis of reconfigurable biomolecular materials. Now, the combination of dynamic and structural DNA nanotechnology has enabled the dynamic control of the assembly and disassembly of DNA nanotubes. The process involves minimal synthetic gene systems, including an autonomous molecular oscillator.

[1]  Y. Rondelez Competition for catalytic resources alters biological network dynamics. , 2012, Physical review letters.

[2]  Erik Winfree,et al.  Determining hydrodynamic forces in bursting bubbles using DNA nanotube mechanics , 2015, Proceedings of the National Academy of Sciences.

[3]  Xiangling Xiong,et al.  Using azobenzene incorporated DNA aptamers to probe molecular binding interactions. , 2011, Bioconjugate chemistry.

[4]  Lulu Qian,et al.  Supporting Online Material Materials and Methods Figs. S1 to S6 Tables S1 to S4 References and Notes Scaling up Digital Circuit Computation with Dna Strand Displacement Cascades , 2022 .

[5]  E. Winfree,et al.  Construction of an in vitro bistable circuit from synthetic transcriptional switches , 2006, Molecular systems biology.

[6]  R. Wade,et al.  Characterization of microtubule protofilament numbers. How does the surface lattice accommodate? , 1990, Journal of molecular biology.

[7]  Deepak Mishra,et al.  A load driver device for engineering modularity in biological networks , 2014, Nature Biotechnology.

[8]  Pamela E. Constantinou,et al.  From Molecular to Macroscopic via the Rational Design of a Self-Assembled 3D DNA Crystal , 2009, Nature.

[9]  Eduardo Sontag,et al.  Modular cell biology: retroactivity and insulation , 2008, Molecular systems biology.

[10]  Franco Blanchini,et al.  Molecular Titration Promotes Oscillations and Bistability in Minimal Network Models with Monomeric Regulators. , 2016, ACS synthetic biology.

[11]  Ximin He,et al.  Synthetic homeostatic materials with chemo-mechano-chemical self-regulation , 2012, Nature.

[12]  Jongmin Kim,et al.  In vitro synthetic transcriptional networks , 2007 .

[13]  D. Chrétien,et al.  New data on the microtubule surface lattice , 1991, Biology of the cell.

[14]  N. Seeman,et al.  Programmable materials and the nature of the DNA bond , 2015, Science.

[15]  Luvena L. Ong,et al.  Three-Dimensional Structures Self-Assembled from DNA Bricks , 2012, Science.

[16]  Conrad Steenberg,et al.  NUPACK: Analysis and design of nucleic acid systems , 2011, J. Comput. Chem..

[17]  J. Reif,et al.  A two-state DNA lattice switched by DNA nanoactuator. , 2003, Angewandte Chemie.

[18]  Sivaraj Sivaramakrishnan,et al.  Engineering Circular Gliding of Actin Filaments Along Myosin-Patterned DNA Nanotube Rings To Study Long-Term Actin-Myosin Behaviors. , 2016, ACS nano.

[19]  Pekka Orponen,et al.  DNA rendering of polyhedral meshes at the nanoscale , 2015, Nature.

[20]  Cecilia Conde,et al.  Microtubule assembly, organization and dynamics in axons and dendrites , 2009, Nature Reviews Neuroscience.

[21]  Hao Yan,et al.  Structural DNA Nanotechnology: State of the Art and Future Perspective , 2014, Journal of the American Chemical Society.

[22]  Faisal A. Aldaye,et al.  Organization of Intracellular Reactions with Rationally Designed RNA Assemblies , 2011, Science.

[23]  Y. Sakai,et al.  Programming an in vitro DNA oscillator using a molecular networking strategy , 2011, Molecular systems biology.

[24]  T. Mitchison,et al.  Microtubule polymerization dynamics. , 1997, Annual review of cell and developmental biology.

[25]  N. Seeman,et al.  Design and self-assembly of two-dimensional DNA crystals , 1998, Nature.

[26]  Shawn M. Douglas,et al.  Self-assembly of DNA into nanoscale three-dimensional shapes , 2009, Nature.

[27]  D. Bartel,et al.  Synthesizing life : Paths to unforeseeable science & technology , 2001 .

[28]  Xi Chen,et al.  Rational, modular adaptation of enzyme-free DNA circuits to multiple detection methods , 2011, Nucleic acids research.

[29]  Adam H. Marblestone,et al.  Rapid prototyping of 3D DNA-origami shapes with caDNAno , 2009, Nucleic acids research.

[30]  Harold Fellermann,et al.  Specific and reversible DNA-directed self-assembly of oil-in-water emulsion droplets , 2012, Proceedings of the National Academy of Sciences.

[31]  E. Winfree,et al.  Synthetic in vitro transcriptional oscillators , 2011, Molecular systems biology.

[32]  J. Reif,et al.  DNA nanotubes self-assembled from triple-crossover tiles as templates for conductive nanowires. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[33]  Hao Yan,et al.  Complex wireframe DNA origami nanostructures with multi-arm junction vertices. , 2015, Nature nanotechnology.

[34]  G. Gundersen,et al.  Beyond polymer polarity: how the cytoskeleton builds a polarized cell , 2008, Nature Reviews Molecular Cell Biology.

[35]  E. Winfree,et al.  Diversity in the dynamical behaviour of a compartmentalized programmable biochemical oscillator. , 2014, Nature chemistry.

[36]  P. Iglesias,et al.  An Excitable Signal Integrator Couples to an Idling Cytoskeletal Oscillator to Drive Cell Migration , 2013, Nature Cell Biology.

[37]  Erik Winfree,et al.  Neural Network Computation by In Vitro Transcriptional Circuits , 2004, NIPS.

[38]  P. Rothemund,et al.  Programmable molecular recognition based on the geometry of DNA nanostructures. , 2011, Nature chemistry.

[39]  Hao Yan,et al.  Organizing DNA origami tiles into larger structures using preformed scaffold frames. , 2011, Nano letters.

[40]  Eun Jeong Cho,et al.  Applications of aptamers as sensors. , 2009, Annual review of analytical chemistry.

[41]  P. Mattila,et al.  Filopodia: molecular architecture and cellular functions , 2008, Nature Reviews Molecular Cell Biology.

[42]  E. Winfree,et al.  Design and characterization of programmable DNA nanotubes. , 2004, Journal of the American Chemical Society.

[43]  J. Reif,et al.  DNA-Templated Self-Assembly of Protein Arrays and Highly Conductive Nanowires , 2003, Science.

[44]  Sahand Jamal Rahi,et al.  The CDK-APC/C Oscillator Predominantly Entrains Periodic Cell-Cycle Transcription , 2016, Cell.

[45]  Erik Winfree,et al.  Integrating DNA strand-displacement circuitry with DNA tile self-assembly , 2013, Nature Communications.

[46]  Faisal A. Aldaye,et al.  Dynamic DNA templates for discrete gold nanoparticle assemblies: control of geometry, modularity, write/erase and structural switching. , 2007, Journal of the American Chemical Society.

[47]  F. Ricci,et al.  pH-Controlled Assembly of DNA Tiles , 2016, Journal of the American Chemical Society.

[48]  Teruo Fujii,et al.  Bottom-up construction of in vitro switchable memories , 2012, Proceedings of the National Academy of Sciences.

[49]  H. Dietz,et al.  Dynamic DNA devices and assemblies formed by shape-complementary, non–base pairing 3D components , 2015, Science.

[50]  Erik Winfree,et al.  Enzyme-free nucleic acid dynamical systems , 2017, Science.

[51]  N. Seeman,et al.  DNA double-crossover molecules. , 1993, Biochemistry.

[52]  Axel Ekani-Nkodo,et al.  Joining and scission in the self-assembly of nanotubes from DNA tiles. , 2004, Physical review letters.

[53]  Michael Blank,et al.  Aptamer Selection Technology and Recent Advances , 2015, Molecular therapy. Nucleic acids.

[54]  Hari K. K. Subramanian,et al.  pH-Driven Reversible Self-Assembly of Micron-Scale DNA Scaffolds. , 2017, Nano letters.

[55]  G. Seelig,et al.  Dynamic DNA nanotechnology using strand-displacement reactions. , 2011, Nature chemistry.

[56]  Jongmin Kim,et al.  A coarse-grained model captures the temporal evolution of DNA nanotube length distributions , 2018, Natural Computing.

[57]  Rui Gan,et al.  Cell-free protein synthesis: applications come of age. , 2012, Biotechnology advances.

[58]  Matthew N. O’Brien,et al.  Anisotropic nanoparticle complementarity in DNA-mediated co-crystallization. , 2015, Nature materials.

[59]  P. Johnson Thermodynamics of the Polymerization of Protein , 1976 .

[60]  G. Seelig,et al.  Enzyme-Free Nucleic Acid Logic Circuits , 2022 .

[61]  Luca Cardelli,et al.  Programmable chemical controllers made from DNA. , 2013, Nature nanotechnology.

[62]  J. Reif,et al.  Construction, analysis, ligation, and self-assembly of DNA triple crossover complexes , 2000 .

[63]  Hao Yan,et al.  Control of Self-Assembly of DNA Tubules Through Integration of Gold Nanoparticles , 2009, Science.

[64]  Rebecca Schulman,et al.  Directing self-assembly of DNA nanotubes using programmable seeds. , 2013, Nano letters.

[65]  Elisa Franco,et al.  T7 RNA polymerase non-specifically transcribes and induces disassembly of DNA nanostructures , 2018, Nucleic acids research.

[66]  P. Rothemund Folding DNA to create nanoscale shapes and patterns , 2006, Nature.

[67]  Erik Winfree,et al.  Robustness and modularity properties of a non-covalent DNA catalytic reaction , 2010, Nucleic acids research.

[68]  J. Howard,et al.  Flexural rigidity of microtubules and actin filaments measured from thermal fluctuations in shape , 1993, The Journal of cell biology.

[69]  Anna Gutowska,et al.  Lessons from nature: stimuli-responsive polymers and their biomedical applications. , 2002, Trends in biotechnology.

[70]  D. Bartel,et al.  Synthesizing life , 2001, Nature.

[71]  Erik Winfree,et al.  Diversity in the dynamical behaviour of a compartmentalized programmable biochemical oscillator , 2014 .

[72]  R. Murray,et al.  Timing molecular motion and production with a synthetic transcriptional clock , 2011, Proceedings of the National Academy of Sciences.

[73]  Charles Boone,et al.  Formins: signaling effectors for assembly and polarization of actin filaments , 2003, Journal of Cell Science.

[74]  Erik Winfree,et al.  Direct atomic force microscopy observation of DNA tile crystal growth at the single-molecule level. , 2012, Journal of the American Chemical Society.

[75]  Stephen Mann,et al.  Life as a nanoscale phenomenon. , 2008, Angewandte Chemie.

[76]  E. Winfree,et al.  Synthesis of crystals with a programmable kinetic barrier to nucleation , 2007, Proceedings of the National Academy of Sciences.

[77]  Jongmin Kim,et al.  A Coarse-Grained Model of DNA Nanotube Population Growth , 2016, DNA.

[78]  Bernard Yurke,et al.  Using DNA to Power Nanostructures , 2003, Genetic Programming and Evolvable Machines.

[79]  G. Seelig,et al.  DNA as a universal substrate for chemical kinetics , 2010, Proceedings of the National Academy of Sciences.