A microRNA-initiated DNAzyme motor operating in living cells

Synthetic DNA motors have great potential to mimic natural protein motors in cells but the operation of synthetic DNA motors in living cells remains challenging and has not been demonstrated. Here we report a DNAzyme motor that operates in living cells in response to a specific intracellular target. The whole motor system is constructed on a 20 nm gold nanoparticle (AuNP) decorated with hundreds of substrate strands serving as DNA tracks and dozens of DNAzyme molecules each silenced by a locking strand. Intracellular interaction of a target molecule with the motor system initiates the autonomous walking of the motor on the AuNP. An example DNAzyme motor responsive to a specific microRNA enables amplified detection of the specific microRNA in individual cancer cells. Activated by specific intracellular targets, these self-powered DNAzyme motors will have diverse applications in the control and modulation of biological functions.

[1]  William O. Hancock,et al.  Bidirectional cargo transport: moving beyond tug of war , 2014, Nature Reviews Molecular Cell Biology.

[2]  M. Maguire,et al.  Magnesium chemistry and biochemistry , 2002, Biometals.

[3]  Moss Y. Zhao,et al.  Disease-Specific Target Gene Expression Profiling of Molecular Imaging Probes: Database Development and Clinical Validation , 2014, Molecular imaging.

[4]  A. Turberfield,et al.  A DNA-fuelled molecular machine made of DNA , 2022 .

[5]  F. Wolf,et al.  Cell (patho)physiology of magnesium. , 2008, Clinical science.

[6]  Robert M. Dirks,et al.  An autonomous polymerization motor powered by DNA hybridization , 2007, Nature Nanotechnology.

[7]  Maode Lai,et al.  Binding-Induced DNA Nanomachines Triggered by Proteins and Nucleic Acids. , 2015, Angewandte Chemie.

[8]  Chengde Mao,et al.  An autonomous DNA nanomotor powered by a DNA enzyme. , 2004, Angewandte Chemie.

[9]  Ronald D Vale,et al.  The Molecular Motor Toolbox for Intracellular Transport , 2003, Cell.

[10]  Yang Liu,et al.  High-speed DNA-based rolling motors powered by RNase H , 2015, Nature nanotechnology.

[11]  A. Turberfield,et al.  Direct observation of stepwise movement of a synthetic molecular transporter. , 2011, Nature nanotechnology.

[12]  Yi Lu,et al.  Photocaged DNAzymes as a general method for sensing metal ions in living cells. , 2014, Angewandte Chemie.

[13]  N. Seeman,et al.  A precisely controlled DNA biped walking device , 2004 .

[14]  Tae-Gon Cha,et al.  Optical nanosensor architecture for cell-signaling molecules using DNA aptamer-coated carbon nanotubes. , 2011, ACS nano.

[15]  Jonathan Bath,et al.  A DNA-based molecular motor that can navigate a network of tracks. , 2012, Nature nanotechnology.

[16]  Wenhu Zhou,et al.  DNAzyme hybridization, cleavage, degradation, and sensing in undiluted human blood serum. , 2015, Analytical chemistry.

[17]  G. F. Joyce,et al.  Mechanism and utility of an RNA-cleaving DNA enzyme. , 1998, Biochemistry.

[18]  Y. Bignon,et al.  Identification of miR-10b, miR-26a, miR-146a and miR-153 as potential triple-negative breast cancer biomarkers , 2015, Cellular Oncology.

[19]  Jing Pan,et al.  Design Principles of DNA Enzyme-Based Walkers: Translocation Kinetics and Photoregulation. , 2015, Journal of the American Chemical Society.

[20]  A. Ellington,et al.  A stochastic DNA walker that traverses a microparticle surface , 2015, Nature nanotechnology.

[21]  B. Bay,et al.  Gold nanoparticles in cancer therapy , 2011, Acta Pharmacologica Sinica.

[22]  David R. Liu,et al.  Autonomous Multistep Organic Synthesis in a Single Isothermal Solution Mediated by a DNA Walker , 2010, Nature nanotechnology.

[23]  P. Yin,et al.  A DNAzyme that walks processively and autonomously along a one-dimensional track. , 2005, Angewandte Chemie.

[24]  Peng Yin,et al.  Optimizing the specificity of nucleic acid hybridization. , 2012, Nature chemistry.

[25]  Amber L. Wells,et al.  Myosin VI is a processive motor with a large step size , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[26]  J. Reif,et al.  A unidirectional DNA walker that moves autonomously along a track. , 2004, Angewandte Chemie.

[27]  Nikolai G Khlebtsov,et al.  Uptake of engineered gold nanoparticles into mammalian cells. , 2014, Chemical reviews.

[28]  Itamar Willner,et al.  DNA machines: bipedal walker and stepper. , 2011, Nano letters.

[29]  Sarit S. Agasti,et al.  Recognition-Mediated Activation of Therapeutic Gold Nanoparticles Inside Living Cells , 2010, Nature chemistry.

[30]  C. Mirkin,et al.  Oligonucleotide loading determines cellular uptake of DNA-modified gold nanoparticles. , 2007, Nano letters.

[31]  N. Pierce,et al.  A synthetic DNA walker for molecular transport. , 2004, Journal of the American Chemical Society.

[32]  Junbo Chen,et al.  Enzyme-Powered Three-Dimensional DNA Nanomachine for DNA Walking, Payload Release, and Biosensing. , 2016, ACS nano.

[33]  R. Weinberg,et al.  Tumour invasion and metastasis initiated by microRNA-10b in breast cancer , 2007, Nature.

[34]  Na Liu,et al.  A plasmonic nanorod that walks on DNA origami , 2015, Nature Communications.

[35]  Jing Pan,et al.  A synthetic DNA motor that transports nanoparticles along carbon nanotubes. , 2014, Nature nanotechnology.

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

[37]  Chad A Mirkin,et al.  Nano-flares: probes for transfection and mRNA detection in living cells. , 2007, Journal of the American Chemical Society.

[38]  Aamir Ahmad,et al.  The Role of MicroRNAs in Breast Cancer Migration, Invasion and Metastasis , 2012, International journal of molecular sciences.

[39]  Weihong Tan,et al.  An autonomous and controllable light-driven DNA walking device. , 2012, Angewandte Chemie.

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

[41]  Eric L. Null,et al.  Metal ion as both a cofactor and a probe of metal-binding sites in a uranyl-specific DNAzyme: a uranyl photocleavage study , 2013, Nucleic acids research.

[42]  H. Cheng,et al.  Noninvasive Manganese-Enhanced Magnetic Resonance Imaging for Early Detection of Breast Cancer Metastatic Potential , 2014, Molecular imaging.

[43]  N. Hirokawa,et al.  Kinesin superfamily motor proteins and intracellular transport , 2009, Nature Reviews Molecular Cell Biology.

[44]  Erik Winfree,et al.  Molecular robots guided by prescriptive landscapes , 2010, Nature.

[45]  Richard A. Muscat,et al.  A programmable molecular robot. , 2011, Nano letters.

[46]  Yi Lu,et al.  A lead-dependent DNAzyme with a two-step mechanism. , 2003, Biochemistry.

[47]  R. Vale,et al.  Kinesin Walks Hand-Over-Hand , 2004, Science.

[48]  N. Seeman,et al.  A Proximity-Based Programmable DNA Nanoscale Assembly Line , 2010, Nature.

[49]  Robert H. Singer,et al.  In the right place at the right time: visualizing and understanding mRNA localization , 2014, Nature Reviews Molecular Cell Biology.

[50]  Jing Pan,et al.  Recent progress on DNA based walkers. , 2015, Current opinion in biotechnology.

[51]  Richard A. Muscat,et al.  DNA nanotechnology from the test tube to the cell. , 2015, Nature nanotechnology.

[52]  A. Turberfield,et al.  A free-running DNA motor powered by a nicking enzyme. , 2005, Angewandte Chemie.