US 11,326,196 B2Granted

System and method for synthesis of DNA particles and use thereof

AssigneesVilnius University, Eth 7Lirich

InventorsLinas Mazutis, Greta Stonyte, Vaidotas Kiseliovas, Rapolas Zilionis, Arvydas Janulaitis, Robertas Galinis, Sabine Studer, Donald Hilvert

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Abstract

Disclosed is a system and method for production of DNA particles and use thereof. The DNA particles can be produced by amplification of nucleic acid molecule(s). Alternatively, DNA particles can be prepared by condensing multiple DNA molecules. The DNA condensation into a particle is mainly triggered by pyrophosphate and positively charged cations (e.g. magnesium). DNA particles can be applied for numerous biological applications but not limited to directed evolution, proteomics, drug delivery and imaging. DNA particles can be used to synthesize proteins using in vitro transcription/translation reaction.

Claims10

Dependent claims

A method of generating individual particles each using a single nucleic acid molecule template by means of enzymatic amplification in aqueous micro-droplets, wherein each particle comprises nucleic acid(s), pyrophosphate(s), and metal ions, wherein said method of producing the individual particles comprises:

(i) providing one or more nucleic acid molecule templates to be amplified;
(ii) providing nucleic acid amplification reaction ingredients including polymerase, primers, dNTP, salts, non-ionic detergent(s);
(iii) partitioning nucleic acid molecules and amplification reaction ingredients in the aqueous micro-droplets using a microfluidics system such that one aqueous micro-droplet contains, on average, no more than one nucleic acid molecule template;
(iv) incubating for a given period of time to allow nucleic acid amplification and formation of the particles to occur;
and
(v) releasing particles from the aqueous micro-droplets.

3 more elements

Dependencies are inferred from the claim text.

Facts

Application: 16/069,404Filed 10 Jan 2017 · us provisional application
Priority: 11 Jan 2016
Published: US 2019/0002943 A13 Jan 2019
Granted: 10 May 2022
Term ends: Not estimatedThe source records don’t support an estimate.

Classification12 codes

C12P 19/34
Nucleotides: polynucleotides, e.g. nucleic acids, oligoribonucleotides
A61K 9/50
Preparations in capsules, e.g. of gelatin, of chocolate: microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets…
B01J 19/0093
Chemical, physical or physico-chemical processes in general; Their relevant apparatus: microreactors, e.g. miniaturised or microfabricated reactors
B01L 3/502784
Characterised by multiphase flow arrangements: specially adapted for droplet or plug flow, e.g. digital microfluidics