title: DNA Nanoparticles for Improved Protein Synthesis In Vitro.
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.
authors.affiliation: Institute of Biotechnology Vilnius University, 8 Graiciuno street, 02241, Vilnius, Lithuania.
authors.forename: Robertas
authors.initials: R
authors.lastname: Galinis
authors.affiliation: Institute of Biotechnology Vilnius University, 8 Graiciuno street, 02241, Vilnius, Lithuania.
authors.forename: Greta
authors.initials: G
authors.lastname: Stonyte
authors.affiliation: Institute of Biotechnology Vilnius University, 8 Graiciuno street, 02241, Vilnius, Lithuania.
authors.forename: Vaidotas
authors.initials: V
authors.lastname: Kiseliovas
authors.affiliation: Institute of Biotechnology Vilnius University, 8 Graiciuno street, 02241, Vilnius, Lithuania.
authors.forename: Rapolas
authors.initials: R
authors.lastname: Zilionis
authors.affiliation: Laboratory of Organic Chemistry, ETH Zurich, Vladimir-Prelog-Weg 1-5, 8093, Zurich, Switzerland.
authors.forename: Sabine
authors.initials: S
authors.lastname: Studer
authors.affiliation: Laboratory of Organic Chemistry, ETH Zurich, Vladimir-Prelog-Weg 1-5, 8093, Zurich, Switzerland.
authors.forename: Donald
authors.initials: D
authors.lastname: Hilvert
authors.affiliation: Institute of Biotechnology Vilnius University, 8 Graiciuno street, 02241, Vilnius, Lithuania.
authors.forename: Arvydas
authors.initials: A
authors.lastname: Janulaitis
authors.affiliation: Institute of Biotechnology Vilnius University, 8 Graiciuno street, 02241, Vilnius, Lithuania. linas.mazutis@bti.vu.lt.
authors.forename: Linas
authors.initials: L
authors.lastname: Mazutis
mesh_terms: D004247:DNA / Q000737:chemistry*; D005453:Fluorescence; D056656:Lab-On-A-Chip Devices; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles; D011506:Proteins / Q000138:chemical synthesis*
chemical_list: D011506:Proteins; D004247:DNA
keywords: DNA; microfluidics; nanoparticles; protein expression; pyrophosphate
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