title: Genetic encoding of DNA nanostructures and their self-assembly in living bacteria.
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 μm(2)) 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.
authors.affiliation: Synthetic Biology Center, Department of Biological Engineering, Massachusetts Institute of Technology, 500 Technology Square NE47-140, Cambridge, Massachusetts 02139, USA.
authors.forename: Johann
authors.initials: J
authors.lastname: Elbaz
authors.affiliation: Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA.|Department of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Peng
authors.initials: P
authors.lastname: Yin
authors.affiliation: Synthetic Biology Center, Department of Biological Engineering, Massachusetts Institute of Technology, 500 Technology Square NE47-140, Cambridge, Massachusetts 02139, USA.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Voigt
mesh_terms: D020029:Base Pairing; D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D054303:HIV Reverse Transcriptase / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D018625:Microscopy, Atomic Force; D008957:Models, Genetic; D008967:Molecular Biology / Q000379:methods; D008969:Molecular Sequence Data; D049329:Nanostructures / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods; D009690:Nucleic Acid Conformation; D017422:Sequence Analysis, DNA
chemical_list: D004277:DNA, Single-Stranded; D008164:Luminescent Proteins; D054303:HIV Reverse Transcriptase