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Genetic encoding of DNA nanostructures and their self-assembly in living bacteria

Author

Listed:
  • Johann Elbaz

    (Synthetic Biology Center, Massachusetts Institute of Technology, 500 Technology Square NE47-140)

  • Peng Yin

    (Wyss Institute for Biologically Inspired Engineering, Harvard University
    Harvard Medical School)

  • Christopher A. Voigt

    (Synthetic Biology Center, Massachusetts Institute of Technology, 500 Technology Square NE47-140)

Abstract

The field of DNA nanotechnology has harnessed the programmability of DNA base pairing to direct single-stranded DNAs (ssDNAs) to assemble into desired 3D structures. Here, we show the ability to express ssDNAs in Escherichia coli (32–205 nt), which can form structures in vivo or be purified for in vitro assembly. Each ssDNA is encoded by a gene that is transcribed into non-coding RNA containing a 3′-hairpin (HTBS). HTBS recruits HIV reverse transcriptase, which nucleates DNA synthesis and is aided in elongation by murine leukemia reverse transcriptase. Purified ssDNA that is produced in vivo is used to assemble large 1D wires (300 nm) and 2D sheets (5.8 μm2) in vitro. Intracellular assembly is demonstrated using a four-ssDNA crossover nanostructure that recruits split YFP when properly assembled. Genetically encoding DNA nanostructures provides a route for their production as well as applications in living cells.

Suggested Citation

  • Johann Elbaz & Peng Yin & Christopher A. Voigt, 2016. "Genetic encoding of DNA nanostructures and their self-assembly in living bacteria," Nature Communications, Nature, vol. 7(1), pages 1-11, September.
    • Handle: RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms11179
    DOI: 10.1038/ncomms11179
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