The present invention is directed to methods and kits for template-free enzymatic synthesis of polynucleotides that include or enable a step of efficiently cleaving the polynucleotide products from its initiator using an endonuclease V activity and initiator with a 3′-penultimate deoxyinosine.
C12Q 1/68 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismesCompositions à cet effetProcédés pour préparer ces compositions faisant intervenir des acides nucléiques
C12P 19/14 - Préparation de composés contenant des radicaux saccharide préparés par action d'une carbohydrase, p. ex. par action de l'alpha-amylase
C12P 19/34 - Polynucléotides, p. ex. acides nucléiques, oligoribonucléotides
2.
Terminal deoxynucleotidyl transferase variants and uses thereof
The present invention is directed to terminal deoxynucleotidyltransferase (TdT) variants that (i) comprise an amino acid sequence that is at least a specified percent identical to an indicated SEQ ID NOs and have at least one substitution at Q455 or at least Q455 plus at least one further substitution at G186, S248, T331, Q390, K394 or H466 (where positions are with respect to SEQ ID NO 1 and functionally equivalent positions in indicated SEQ ID NOs), (ii) are capable of template-free extension of a polynucleotide, and (iii) exhibit enhanced stability or enhanced efficiency in incorporating 3′-0-blocked nucleoside triphosphates into a polynucleotide. The invention is also directed to the use of these TdT variants for synthesizing polynucleotides of any predetermined sequence.
The present invention is directed to methods and kits for template-free enzymatic synthesis of polynucleotides that include or enable a step of efficiently cleaving the polynucleotide products from its initiator using an endonuclease V activity and initiator with a 3′-penultimate deoxyinosine.
C12Q 1/68 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismesCompositions à cet effetProcédés pour préparer ces compositions faisant intervenir des acides nucléiques
C12N 9/12 - Transférases (2.) transférant des groupes contenant du phosphore, p. ex. kinases (2.7)
C12Q 1/6806 - Préparation d’acides nucléiques pour analyse, p. ex. pour test de réaction en chaîne par polymérase [PCR]
C12Q 1/6876 - Produits d’acides nucléiques utilisés dans l’analyse d’acides nucléiques, p. ex. amorces ou sondes
C12N 15/10 - Procédés pour l'isolement, la préparation ou la purification d'ADN ou d'ARN
C12P 19/14 - Préparation de composés contenant des radicaux saccharide préparés par action d'une carbohydrase, p. ex. par action de l'alpha-amylase
The invention relates to a process for preparing a 3′-O-amino-ribonucleotide. It also relates to a compound of formula (III), and its use as a precursor for the synthesis of a 3′-O-amino-ribonucleotide.
In formula (III), B is a nitrogenous base or a protected derivative thereof, G and G′ are identical or different, and are a protecting group, and RN— is a phthalimido or an imino group.
Methods for preparing 3′-O-amino-2′-deoxyribonucleoside-5′-triphosphates with reduced 3′-hydroxy-2′-deoxyribonucleoside-5′-triphosphate contamination by converting 3′-(N-acetone-oxime)-2′-deoxynucleoside triphosphate to 3′-O-amine-2′-deoxynucleoside triphosphate by treatment with an aryl-oxyamine and compositions produced therefrom.
The present invention is directed to methods and kits for template-free enzymatic synthesis of polynucleotides that include or enable a step of efficiently cleaving the polynucleotide products from its initiator using endonuclease V activity and initiator with a 3′-penultimate deoxyinosine.
C12Q 1/68 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismesCompositions à cet effetProcédés pour préparer ces compositions faisant intervenir des acides nucléiques
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.
C12N 15/10 - Procédés pour l'isolement, la préparation ou la purification d'ADN ou d'ARN
C12N 15/11 - Fragments d'ADN ou d'ARNLeurs formes modifiées
C12Q 1/44 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismesCompositions à cet effetProcédés pour préparer ces compositions faisant intervenir une hydrolase une estérase
C40B 50/06 - Procédés biochimiques, p. ex. utilisant des enzymes ou des micro-organismes viables entiers
C40B 50/08 - Synthèse en phase liquide, c.-à-d. dans laquelle tous les blocs servant à créer la bibliothèque sont en phase liquide ou en solution au cours de la création de la bibliothèqueProcédés particuliers de clivage à partir du support liquide
C40B 50/14 - Synthèse en phase solide, c.-à-d. dans laquelle au moins un bloc servant à créer la bibliothèque est lié à un support solide au cours de la création de la bibliothèqueProcédés particuliers de clivage à partir du support solide
C40B 70/00 - Étiquettes ["tags"] ou marqueurs ["labels"] spécialement adaptés à la chimie combinatoire ou aux chimiothèques, p. ex. "tags" fluorescents ou codes-barres
8.
Direct oligonucleotide synthesis on cells and biomolecules
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.
C12N 15/10 - Procédés pour l'isolement, la préparation ou la purification d'ADN ou d'ARN
C12N 15/11 - Fragments d'ADN ou d'ARNLeurs formes modifiées
C12Q 1/44 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismesCompositions à cet effetProcédés pour préparer ces compositions faisant intervenir une hydrolase une estérase
C40B 50/06 - Procédés biochimiques, p. ex. utilisant des enzymes ou des micro-organismes viables entiers
C40B 50/08 - Synthèse en phase liquide, c.-à-d. dans laquelle tous les blocs servant à créer la bibliothèque sont en phase liquide ou en solution au cours de la création de la bibliothèqueProcédés particuliers de clivage à partir du support liquide
C40B 50/14 - Synthèse en phase solide, c.-à-d. dans laquelle au moins un bloc servant à créer la bibliothèque est lié à un support solide au cours de la création de la bibliothèqueProcédés particuliers de clivage à partir du support solide
C40B 70/00 - Étiquettes ["tags"] ou marqueurs ["labels"] spécialement adaptés à la chimie combinatoire ou aux chimiothèques, p. ex. "tags" fluorescents ou codes-barres
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
09 - Appareils et instruments scientifiques et électriques
Produits et services
Chemicals, reagents, enzymes and reagent kits comprising
enzymes, monomers, solid supports, and synthesis plates, all
for scientific research use, namely, for use in synthesizing
polynucleotides, namely, for amplifying, assembling, and
modifying genetic fragments, genotyping, modifying genes,
and analyzing biological cells, samples, and systems, in
biological, medical, and clinical laboratories and
facilities using or storing nucleic acid molecules. Chemical reagents and enzymes for medical use, namely, for
use in synthesizing polynucleotides, namely, for amplifying,
assembling, and modifying genetic fragments, genotyping,
modifying genes, and analyzing biological cells, samples,
and systems, in biological, medical, and clinical
laboratories and facilities using or storing nucleic acid
molecules. Apparatus and instruments for synthesizing polynucleotides,
namely, DNA and RNA, and parts and accessories therefor,
namely, bottles, cartridges, vials, reaction plates,
synthesis supports, reaction tubes, and related computer
hardware and downloadable computer software for use in
operating and interfacing with apparatus and instruments for
synthesizing polynucleotides.
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
09 - Appareils et instruments scientifiques et électriques
Produits et services
(1) Chemicals, reagents, enzymes and reagent kits comprising enzymes, monomers, solid supports, and synthesis plates, all for scientific research use, namely, for use in synthesizing polynucleotides, namely, for amplifying, assembling, and modifying genetic fragments, genotyping, modifying genes, and analyzing biological cells, samples, and systems, in biological, medical, and clinical laboratories and facilities using or storing nucleic acid molecules.
(2) Chemical reagents and enzymes for medical use, namely for use in polynucleotide synthesis, namely for amplification, assembly, modification of genetic fragments, genotyping, gene modification, and analysis of biological systems, samples, and cells, in clinical, medical and biological installations and laboratories using or storing nucleic acid molecules, excluding any products directly intended for the cardiovascular field.
(3) Instruments, namely, polynucleotide synthesizers for synthesizing polynucleotides, namely, DNA and RNA, and parts and accessories therefor, namely, bottles, cartridges, vials, reaction plates, synthesis supports, reaction tubes, and related computer hardware and downloadable computer software for use in operating and interfacing with instruments, namely, polynucleotide synthesizers for synthesizing polynucleotides.
11.
Variants of terminal deoxynucleotidyl transferase 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.
