131 results about "Oligonucleotide synthesis" patented technology

The present disclosure, among other things, provides technologies for synthesis, including reagents and methods for stereoselective synthesis. In some embodiments, the present disclosure provides compounds useful as chiral auxiliaries. In some embodiments, the present disclosure provides reagents and methods for oligonucleotide synthesis. In some embodiments, the present disclosure provides reagents and methods for chirally controlled preparation of oligonucleotides. In some embodiments, technologies of the present disclosure are particularly useful for constructing challenging internucleotidic linkages, providing high yields and stereoselectivity.

The invention provides a synthesis method and application of nucleoside monomer analogues and oligonucleotides. R on the nucleoside monomer is a basic group; r1 comprises one or more of methoxyl, methoxyethyl, fluorine, hydroxyl or hydrogen; r2 comprises one or more of hydroxyl, monophosphoric acid, diphosphoric acid, triphosphoric acid or thio-modified phosphoric acid; and R3 comprises one or more of methyl, ethyl, propyl or isopropyl. The method can solve the problem of low reversible end capping activity of oligonucleotide in the prior art, and is suitable for the field of oligonucleotide synthesis.

The present disclosure relates to methods of using single-stranded RNA ligase for synthesis of oligonucleotides, including oligonucleotides containing a conjugate moiety. In some embodiments, the present disclosure further provides recombinant single-stranded RNA ligases, polynucleotides encoding the recombinant single-stranded RNA ligases, and compositions of the single-stranded RNA ligases.

The invention discloses an oligonucleotide synthesis process of a sulfo-PMO structure. In the oligonucleotide synthesis process of the sulfo-PMO structure, the molecular structural formula of sulfo-PMO is shown in the specification. According to the invention, the molecular structure of thio-PMO is specifically limited, and the synthesis of nucleotide phosphate amide and (thio) phosphoric acid amide in the solid-phase synthesis oligonucleotide is carried out along the direction from 5'to 3 ', that is, in each cycle period, the O5'or O6' position of the nucleotide phosphate amide and (thio) phosphoric acid amide monomer is firstly subjected to coupling reaction, and then the protecting group is removed at the O3 'or N3' position, so that the oligonucleotide phosphate amide and (thio) phosphoric acid amide are obtained. And releasing active hydroxyl or amino / amido to enter the next cycle. The protecting group adopted by the sulfo-PMO molecule is consistent with the protecting groups of other nucleotides, so that the conversion rate of each step of coupling reaction can be conveniently detected on line.

The present disclosure, among other things, provides technologies for oligonucleotide synthesis. In some embodiments, the present disclosure provides phosphoramidites and methods for synthesis thereof. In some embodiments, provided methods provides higher yields and / or purities. In some embodiments, provided methods remove byproducts without contact with an aqueous solution.

The application provides a method for inkjet printing of oligonucleotides, and belongs to the technical field of in-situ synthesis of oligonucleotides.The method comprises the following steps: modifying the surface of a solid carrier with a chemically active functional group; depositing oligonucleotide synthesis reagents onto the surface of the modified solid carrier by inkjet to perform in-situ synthesis; and the solvent in the oligonucleotide synthesis reagents comprises one or two of glutaronitrile, acetonitrile and propylene carbonate.The application uses a new type of difficult-to-vaporize solvent, greatly reduces the volatilization of reagents during inkjet synthesis and the generation of organic vapors during synthesis, and thus maintains a high in-situ synthesis reaction efficiency of oligonucleotides.

The present disclosure provides methods for preparing oligonucleotides. In certain embodiments, the present disclosure provides methods for preparing oligonucleotides using cyclic-phosphate or cyclic-thiophosphate nucleosides. In certain embodiments, the present disclosure provides methods for preparing oligonucleotides using nucleophiles selected from substituted thiol nucleophiles, substituted nitrogen-based nucleophiles, substituted phosphine nucleophiles, halides, pseudohalides, azides, and substituted lithium-based nucleophiles.

The invention belongs to the field of oligonucleotide synthesis, and particularly relates to a polymer-loaded glass carrier, a preparation method and application thereof, and an oligonucleotide synthesis method. The polymer-loaded glass carrier comprises a glass carrier and a modified styrene / nitrogen-containing heterocyclic ring copolymer coating the surface of the glass carrier, the ratio of the thickness of the modified styrene / nitrogen-containing heterocyclic ring copolymer to the average pore size of the glass carrier is 1: (200-250), and the modified styrene / nitrogen-containing heterocyclic ring copolymer has a structure as shown in a formula (1). The preparation method is characterized in that a nitrogen-containing heterocyclic ring is introduced into a polystyrene molecular chain and coupled to CPG, and meanwhile, the ratio of the thickness of the modified styrene / nitrogen-containing heterocyclic ring copolymer to the average pore size of CPG is strictly controlled, so that the effective load of nucleotide can be effectively improved; meanwhile, compared with the method that a nitrogen-containing heterocyclic compound is independently introduced into an oligonucleotide synthesis system, the synthesis efficiency and purity of nucleic acid can be remarkably improved. # imgabs0 #

This invention provides an oligonucleotide synthesis catalyst and its preparation method, as well as a method for synthesizing oligonucleotides. The oligonucleotide synthesis catalyst comprises at least one of a tetrazolium, azole salt, pyridine salt, and imidazole salt compound as the main component, and a polar additive. The oligonucleotide synthesis catalyst of this invention solves the problems of current oligonucleotide synthesis processes, such as the difficulty in synthesizing complex oligonucleotide sequences (e.g., oligonucleotide chains that are too long, have high GC content, or have repetitive or continuous sequences that easily generate secondary structures), the difficulty in obtaining the target oligonucleotide fragment, or the high impurity content and poor quality of the target oligonucleotide fragment. When the oligonucleotide synthesis catalyst of this invention is applied to oligonucleotide synthesis, the success rate of synthesizing high-GC-content oligonucleotides can be increased, and the quality of oligonucleotides can be improved.

The invention relates to the technical field of biological medicine, in particular to an improved preparation method of a GalNAc compound containing a ribose ring. Compared with an existing traditional synthesis route, the method has the advantages that the total yield of the whole reaction is remarkably increased, impurities are reduced, the purification mode is simplified, the cost is reduced, the method is environment-friendly and suitable for industrial production, and the obtained solid-phase carrier with high loading capacity is beneficial to amplified production of downstream oligonucleotide solid-phase synthesis.

This invention provides a high-throughput method for constructing dual sgRNA libraries and its applications. The method utilizes high-throughput microarray synthesis technology to prepare a set of DNA fragments containing multiple dual sgRNA expression cassettes in a single step. After amplification, these fragments are assembled with a target vector containing a first promoter and a second gRNA backbone sequence in a first round of directed assembly to obtain a preliminary recombinant plasmid set. Then, linearized enzyme digestion and homologous recombination technology are used to insert a fragment containing a transcription termination sequence and a complete second promoter to complete the construction of the dual sgRNA expression unit. Finally, transformation and amplification yield the dual sgRNA plasmid library. This invention avoids the high error rate and high cost of long-chain oligonucleotide synthesis by utilizing microarray synthesis and simplifies the operation process through two rounds of directed assembly, significantly improving the throughput, fidelity, and efficiency of library construction. It is applicable to the construction of genome-wide dual sgRNA libraries, providing an efficient and reliable technical platform for high-throughput gene function screening, drug target discovery, and gene interaction research based on CRISPR.

The present disclosure, among other things, provides technologies for synthesis, including reagents and methods for stereoselective synthesis. In some embodiments, the present disclosure provides compounds useful as chiral auxiliaries. In some embodiments, the present disclosure provides reagents and methods for oligonucleotide synthesis. In some embodiments, the present disclosure provides reagents and methods for chirally controlled preparation of oligonucleotides. In some embodiments, technologies of the present disclosure are particularly useful for constructing challenging internucleotidic linkages, providing high yields and stereoselectivity.

An improved protocol for the synthesis of oligonucleotides is provided. A liquid composition for the cleavage of a di(p-methoxyphenyl)phenylmethyl protecting group from a hydroxyl group and the use thereof in chemical synthesis of oligonucleotides is disclosed.

Methods and systems for synthesizing oligonucleotides in a reaction chamber comprising a solid support. Nucleotide monomers are selected for addition to oligonucleotides. An inlet stream comprising selected monomers is fed into a reaction chamber (column) of the reaction chamber and conditions for oligonucleotide synthesis are provided in the reaction chamber. An outlet stream from the reaction chamber may be split and directed to a waste discharge port and / or an analysis station in which the eluate is analyzed by mass spectrometry (MS) to confirm the identity of the monomers fed to the reaction chamber through the inlet. A determination is made as to whether the outlet stream contains the selected nucleotide monomer or derivative thereof based on results by the MS analysis.

The present disclosure relates to methods and compounds for liquid phase oligonucleotide synthesis employing the use of small molecules with lipophilic groups. Methods for making an oligonucleotide by liquid phase oligonucleotide synthesis using the compounds described herein are also provided.

The invention discloses synthesis of 5 '-amino-3-nucleotide phosphamide and application of the 5'-amino-3-nucleotide phosphamide in oligonucleotide. The phosphamide modified oligonucleotide comprises a nucleotide structural unit shown as a formula I or a formula II,..., the formula I; ... formula II; through specific limitation of a nucleotide phosphamide monomer structure, the prepared oligonucleotide contains a phosphamide structural unit; compared with a traditional oligonucleotide molecule, the oligonucleotide molecule with the phosphamide modified nucleotide structure has more excellent effects on pharmaceutical parameters such as enzymatic degradation resistance, stability and the like, and in addition, phosphamide modification can improve the binding performance between the modified oligonucleotide molecule and environmental ions; a novel chemical modification strategy is provided for research, development and application of oligonucleotide drugs.

Apparatuses and a method for plate-based oligonucleotide synthesis are disclosed. In one example, an apparatus used in oligonucleotide synthesis includes a synthesis adapter base to receive a commercially available synthesis plate. A keeper is used to apply pressure to the commercially available synthesis plate, and a sealing element is used to seal the commercially-available synthesis plate to the system adapter base. Other methods and apparatuses are disclosed.

This disclosure provides electrochemically-cleavable linkers with cleavage potentials that are less than the redox potential of the solvent in which the linkers are used. In some applications, the solvent may be water or an aqueous buffer solution. The linkers may be used to link a nucleotide to a bound group. The linkers include a cleavable group which may be one of a methoxybenzyl alcohol, an ester, a propargyl thioether, or a trichloroethyl ether. The linkers may be cleaved in solvent by generating an electrode potential that is less than the redox potential of the solvent. In some implementations, an electrode array may be used to generate localized electrode potentials which selectively cleave linkers bound to the activated electrode. Uses for the linkers include attachment of blocking groups to nucleotides in enzymatic oligonucleotide synthesis.

The purpose of the present disclosure is to provide an amide-crosslinked amidite monomer which contains a guanine base and can contribute to a reduction in the amount of the amidite monomer used during oligonucleotide synthesis. The present disclosure relates to a crystal of an amide-crosslinked compound represented by formula (1). (In the formula, P1 represents a protecting group for a hydroxyl group and P2 represents a protecting group for an amino group.

The invention relates to a composite group oligonucleotide synthesis carrier as well as a preparation method and application thereof. The composite group oligonucleotide synthesis carrier comprises a carrier and groups fixed on the carrier, the groups comprise an activator group and an oligonucleotide group, and the activator group comprises a triazole group and a derivative thereof. According to the invention, the activator group is combined with the activated phosphoramidite group to promote the phosphoramidite group to gather on the carrier, namely, the distance between the activated phosphoramidite group and oligonucleotide is shortened, so that the accuracy of the condensation reaction is improved, the reaction efficiency is improved, and finally, the synthesis efficiency is improved. Meanwhile, in a general nucleic acid synthesis method, in order to ensure full proceeding of the condensation reaction, an excessive amount of monomer reagent is generally needed to be added, and the scheme can greatly improve the accuracy of the condensation reaction, so that less monomer reagent can be allowed to be used, and the cost of the nucleic acid synthesis reaction is reduced.

The present invention provides methods of preparing immobilized engineered terminal nucleotidyl transferase (TnT) polypeptides, and compositions and methods for using these immobilized enzymes in processes for template-independent oligonucleotide synthesis.

This application discloses a method for manufacturing a CPG filter element, comprising: processing a CPG composite filter element, the CPG composite filter element comprising a core block and a CPG matrix integrally formed on the outside of the core block, the core block being resistant to the environment of the oligonucleotide synthesis process and at least partially exposed; and laser marking the exposed portion of the core block to form unique identification information for the CPG composite filter element. In subsequent oligonucleotide solid-phase synthesis operations, the CPG composite filter element of this application can be tracked based on the unique identification information on the core block, and traceability of the synthesis based on the CPG composite filter element can also be achieved. Furthermore, the embodiments of this application avoid the problems of easy detachment, corrosion, or blurring of markings due to traditional marking methods, leading to information loss, and also avoid the destruction of the integrity of the unique identification information during the high-temperature sintering process of the CPG filter element or the destruction of the pore structure of the CPG filter element by high-energy lasers.

The application provides a nucleoside dimer, a preparation method thereof and application in DNA synthesis. The nucleoside dimer of the application adopts a mixed skeleton of methyl phosphate and beta-cyanoethyl phosphoramidite, a new preparation method of the nucleoside dimer is obtained by optimizing a synthesis method and a purification process, and a foundation for high-quality DNA synthesis is laid. On this basis, the nucleoside dimer with the new skeleton is applied to DNA synthesis, an oligonucleotide fragment synthesis method suitable for the dimer is established, and an oligonucleotide fragment with a length of 100 nt is synthesized. Fidelity experiment shows that the error rate of the DNA fragment synthesized by using the dimer is only 3.75 errors / Kb, and high-fidelity DNA synthesis is realized. The preparation method of the nucleoside dimer with the new skeleton can realize large-scale and high-purity preparation, the nucleoside dimer can play an important role in improving the length and fidelity of oligonucleotide synthesis, and has practical value.

The present invention provides a segment for oligonucleotide synthesis that can be more easily purified and supplied in large quantities, a method for producing the same, and a method for synthesizing an oligonucleotide. A segment for oligonucleotide synthesis is synthesized via a phosphoramidite (1) using an amiditizing reagent. TIFF2022065127000032.tif69133

The present invention relates to the technical field of oligonucleotide synthesis. Disclosed are the use of silicon dioxide microspheres in the synthesis of an ultra-long oligonucleotide and a method for synthesizing an ultra-long oligonucleotide, wherein the silicon dioxide microspheres have a specific surface area of less than 1 m2 / g, and preferably, the silicon dioxide microspheres have modification groups on the surface. The method for synthesizing an ultra-long oligonucleotide comprises: performing a coupling reaction on a nucleoside phosphoramidite synthesis unit using the silicon dioxide microspheres having modification groups on the surface as carriers under coupling reaction conditions. The prepared carriers can be used to prepare an ultra-long oligonucleotide.

Apparatuses and a method for plate-based oligonucleotide synthesis are disclosed. In one example, an apparatus used in oligonucleotide synthesis includes a synthesis adapter base to receive a commercially available synthesis plate. A keeper is used to apply pressure to the commercially available synthesis plate, and a sealing element is used to seal the commercially-available synthesis plate to the system adapter base. Other methods and apparatuses are disclosed.

The invention pertains to a method for the synthesis of oligonucleotides using pseudo solid-phase protecting groups, wherein said method comprises a step of subjecting a solution comprising a component (C-0)# to one or more aqueous extractions, wherein the organic phase comprises the component (C-0)#. Said component (C-0)# is a nucleoside or an oligonucleotide, which is covalently bonded to a pseudo solid-phase protecting group and comprises a free backbone hydroxyl group. The aqueous phase of each of said one or more aqueous extractions has a pH-value in the range of 4-7.

Aspects of the disclosure relate to methods and systems for solid phase oligonucleotide synthesis (SPOS). In some embodiments, the methods described by the disclosure comprise systems and methods related to washing after deblocking during solid phase oligonucleotide synthesis. In some embodiments, used deblocking solution is combined with a used washing solution and distilled to provide a distillate wash solution that can be used in a later washing step. In some embodiments, wash solutions are heated or pyridine is added for use during countercurrent washing procedures to increase efficiency. In some embodiments, the methods are combined.

A process of liquid-phase oligonucleotide synthesis (LPOS) is provided which uses the photolabile protecting group 2-(2-nitrophenyl)propyloxycarbonyl (NPPOC). The process may be used for synthesizing an NPPOC-protected oligonucleotide or salt thereof or a precursor thereto. Also provided is the use of NPPOC in liquid phase oligonucleotide synthesis, as well as an NPPOC-protected oligonucleotide or salt thereof which is in solution.

In exemplary aspects, oligonucleotides are synthesized via a microfluidic reaction chamber integrating nanostructure / high-Q dielectric nanoantenna arrays of a metasurface device, the arrays providing synthesis sites. The method comprises; the chamber processing input reagents for producing oligonucleotides at outlets of the chamber; and optically exciting the synthesis sites by altering optical parameter(s) to cause at least one enzymatic synthesis cycle. In more specific examples, the method further includes effecting, after the cycle(s) occurs, PCR amplification of certain of the synthesized products by selecting and optically exciting one or more of the synthesis sites, and / or microfluidically segregating the one or more synthesized products, in response to the amplification step, into homogenous oligo pools.