Dna Script

Institut Pasteur

Dna Script Sas

Centre National De La Recherche Scientifique

Other

National Center for Scientific Research (cnrs)

Thomas Ybert

Elise Champion

Sylvain Gariel

Mikhael Soskine

Marc Delarue

Robert Nicol

Jeffrey Jeddeloh

Xavier Godron

Adrian Horgan

Omar Vivar

Ahmed Said

Sandrine Creton

Gabriel De Crozals

Marc Artigue

Christine Peponnet

Carl Martin

John Luckey

Filip Wojciechowski

Henri Lachaize

Robert Nichol

The invention is directed to methods for massively parallel template-free enzymatic synthesis of a plurality of different polynucleotides of predetermined sequences. In one aspect, methods of the invention employ large scale arrays of reaction sites each associated with at least one working electrode for controlling deprotection and deblocking steps at predetermined user selected sites. In another aspect, the invention provides template-free enzymatic synthesis with proofreading, wherein completed polynucleotides at predetermined reaction sites are sequenced using a sequencing by synthesis technique, particularly employing electrochemically labile blocking groups.

Massively parallel enzymatic synthesis of nucleic acid strands

The invention is directed to systems, apparatus and kits for automated synthesis of a plurality of polynucleotides in an array of reaction chambers using a template-free polymerase. In some embodiments, adaptive elements and processes are provided to monitor and control disruption of the synthesis process and fluid movement by enzyme aggregation.

Systems, apparatus and kits for enzymatic polynucleotide synthesis

The present invention relates to variants of Terminal deoxynucleotidyl Transferase (TdT), each of which (i) has an amino acid sequence similarity to SEQ ID NO: 2, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33 or 35 with corresponding amino acid substitutions, (ii) is capable of synthesizing a nucleic acid fragment without a template and (iii) is capable of incorporating a modified nucleotide into the nucleic acid fragment.

Variants of terminal deoxynucleotidyl transferase and uses thereof

The present invention relates to variant of Family A polymerases able to synthesize a nucleic acid fragment without template and to incorporate a reversible modified terminator nucleotide during the nucleic acid fragment synthesis. The present invention further relates to uses thereof for enzymatic synthesis of nucleic acid molecules.

Variants of family a DNA polymerase and uses thereof

The present invention relates to variants of Terminal deoxynucleotidyl Transferase (TdT), each of which (i) has an amino acid sequence similarity to SEQ ID NO: 2. 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33 or 35 with corresponding amino acid substitutions, (ii) is capable of synthesizing a nucleic acid fragment without a template and (iii) is capable of incorporating a modified nucleotide into the nucleic acid fragment.

The invention is directed to methods for synthesizing oligonucleotides directly on biomolecules or cells living or fixed. In some embodiments, template-free enzymatic synthesis is implemented under biological conditions with successive cycles of (i) enzymatic addition of a 3'-O-blocked nucleoside triphosphate and (ii) enzymatic deblocking of the incorporated nucleotide to regenerate a free 3′ hydroxyl. The invention has applications in single-cell cDNA library construction and analysis.

Methods for extending polynucleotides

The invention relates to a method for synthesising long nucleic acids, including at least one cycle of elongating initial fragments of nucleic acids, including a) a phase comprising the enzymatic addition of nucleotides to said fragments, b) a phase comprising the purification of the fragments having a correct sequence, c) an optional phase of enzymatic amplification, each cycle being performed in a reaction medium which is compatible with enzymatic addition and amplification, such as an aqueous medium, the synthesis method also comprising, at the end of all the elongation cycles, a last step of final amplification. The invention also relates to the use of the method for the production of genes, or sequences of synthetic nucleic acids, DNA or RNA. The invention further relates to a kit for implementing said method.

Method for synthesizing nucleic acids, in particular long nucleic acids, use of said method and kit for implementing said method

Methods for preparing 3'-O-amino-2&#x2032;-deoxyribonucleoside-5&#x2032;-triphosphates with reduced 3&#x2032;-hydroxy-2&#x2032;-deoxyribonucleoside-5&#x2032;-triphosphate contamination by converting 3&#x2032;-(N-acetone-oxime)-2&#x2032;-deoxynucleoside triphosphate to 3&#x2032;-O-amine-2&#x2032;-deoxynucleoside triphosphate by treatment with an aryl-oxyamine and compositions produced therefrom.

Method for preparing 3'-o-amino-2&#x2032;-deoxyribonucleoside-5&#x2032;-triphosphates

The invention is directed to methods for synthesizing oligonucleotides directly on biomolecules or cells living or fixed. In some embodiments, template-free enzymatic synthesis is implemented under biological conditions with successive cycles of (i) enzymatic addition of a 3'-O-blocked nucleoside triphosphate and (ii) enzymatic deblocking of the incorporated nucleotide to regenerate a free 3&#x2032; hydroxyl. The invention has applications in single-cell cDNA library construction and analysis.

Direct oligonucleotide synthesis on cells and biomolecules

A modified nucleotide, intended for the synthesis of long chain nucleic acids by enzymatic processes, comprising a 'natural' nitrogenous base or a natural nitrogenous base analogue, a ribose or deoxyribose carbohydrate, and at least one phosphate group, characterized in that said nucleotide comprises at least one R group, termed the modifier group, carried by said nitrogenous base or analogue and/or by the oxygen in position 3' of the ribose or deoxyribose molecule, making it possible to block the polymerization of said nucleotide and/or to allow the interaction of said nucleotide with another molecule, such as a protein, during the nucleic acid synthesis, R comprising at least one functional terminal group.

Modified nucleotides for synthesis of nucleic acids, a kit containing such nucleotides and their use for the production of synthetic nucleic acid sequences or genes

Growing community of inventors

Paris, France

Thomas Ybert