DNA Nanoparticles for Improved Protein Synthesis In Vitro

Angewandte Chemie
Robertas GalinisLinas Mazutis

Abstract

The amplification and digital quantification of single DNA molecules are important in biomedicine and diagnostics. Beyond quantifying DNA molecules in a sample, the ability to express proteins from the amplified DNA would open even broader applications in synthetic biology, directed evolution, and proteomics. Herein, a microfluidic approach is reported for the production of condensed DNA nanoparticles that can serve as efficient templates for in vitro protein synthesis. Using phi29 DNA polymerase and a multiple displacement amplification reaction, single DNA molecules were converted into DNA nanoparticles containing up to about 10(4)  clonal gene copies of the starting template. DNA nanoparticle formation was triggered by accumulation of inorganic pyrophosphate (produced during DNA synthesis) and magnesium ions from the buffer. Transcription-translation reactions performed in vitro showed that individual DNA nanoparticles can serve as efficient templates for protein synthesis in vitro.

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Citations

Nov 2, 2016·Lab on a Chip·Nan-Nan DengLiang-Yin Chu
Oct 23, 2018·Advanced Materials·Ruihua DongXingyu Jiang
Aug 6, 2019·Advanced Materials·Fengyun LiWeiwei Guo
Jul 26, 2019·Chembiochem : a European Journal of Chemical Biology·Daniel HürtgenVictor Sourjik
Apr 3, 2019·Chembiochem : a European Journal of Chemical Biology·Xiaocui GuoDayong Yang
Sep 25, 2020·Chemistry : a European Journal·Jigang LvDayong Yang
Feb 8, 2018·Current Opinion in Structural Biology·Hans Adrian BunzelDonald Hilvert

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Methods Mentioned

BETA
PCR
chips
MDA
transmission
scanning
atomic
dynamic light scattering
AFM
electrophoresis
Fluorescence

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