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Genetic encoding of DNA nanostructures and their self-assembly in living bacteria
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Journal Article
Genetic encoding of DNA nanostructures and their self-assembly in living bacteria
2016
Overview
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.
DNA nanostructures have the potential to be powerful tools in many areas of biology however they are difficult to manufacture completely
in vivo
. Here the authors combine RNA hairpins and reverse transcription to generate and assemble a complex DNA structure inside the cellular environment.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 14/3
/ 14/35
/ 38
/ 38/22
/ 38/23
/ 38/44
/ 38/47
/ 38/77
/ DNA
/ DNA, Single-Stranded - biosynthesis
/ DNA, Single-Stranded - chemistry
/ DNA, Single-Stranded - genetics
/ E coli
/ HIV Reverse Transcriptase - metabolism
/ Humanities and Social Sciences
/ Leukemia
/ Luminescent Proteins - genetics
/ Luminescent Proteins - metabolism
/ Science
