86 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 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.

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.

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.

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 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

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.

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.

The utility model relates to an oligonucleotide synthesizer, including casing, baroceptor, electrical proportional valve and control subassembly, the casing is provided with the intake cavity, the intake cavity is provided with the intake port and communication port, the electrical proportional valve is provided in the casing, the intake port is used for the communication with the gas source through the electrical proportional valve, and the control subassembly is used for the communication with the gas source through the gas pressure sensor. The machine shell is used for fixing a synthesis container, the communication opening is used for being matched with the synthesis container so that the liquid inlet side of the synthesis container can be communicated with the air inlet cavity and the liquid outlet side of the synthesis container can be communicated with the outside, and the air pressure sensor is arranged on the machine shell and used for detecting the air pressure of the air inlet cavity. The control assembly is electrically connected with the air pressure sensor and the electric proportional valve. The control assembly can control the electric proportional valve so as to adjust the required amount of gas introduced into the gas inlet cavity, so that residual reagents in each reaction step can be completely removed, and the quality of reaction products is ensured.

The invention relates to a liquid-phase synthesis method and application of 5 '-triphosphoric acid modified oligonucleotide. The liquid-phase synthesis method comprises the following steps: synthesis of 5'-terminal phosphoric acid modified oligonucleotide, ammonolysis, salting-out, activation of 5 '-terminal phosphoric acid, connection of pyrophosphoric acid and ethanol precipitation. According to the invention, a liquid phase method is creatively designed to synthesize the oligonucleotide modified by triphosphoric acid, the dependence on a solid phase carrier can be avoided, the synthesis process is safe, the synthesis efficiency is high, the application scene of the synthesis is obviously expanded, and safe, stable and cheap synthesis of the oligonucleotide modified by 5 '-triphosphoric acid is realized.

A reactor system is disclosed. The reactor system includes a vessel configured to contain a solid support, the vessel including: a vessel wall defining a reaction chamber, the reaction chamber having a first end and a second end opposite the first end; a piston operatively arranged at the first end and configured to translate within the reaction chamber; a force measuring device coupled to the piston and configured to measure a load on the piston; a piston driver coupled to the piston; and a processor operably coupled to the force measuring device and the piston driver, the processor configured to: measure a load on the piston using the force measuring device, and adjust a position of the piston using the piston driver based on measuring the load on the piston.

Embodiments for the synthesis of sensitive oligonucleotides as well as insensitive oligonucleotides are provided. Sulfur-based groups are used for the protection of exo-amino groups of nucleobases, phosphate groups and 2′-OH groups, and as cleavable linker for linking oligonucleotides to a support. Oligonucleotide syntheses are achieved under typical conditions using phosphoramidite chemistry with important modifications. To prevent replacing sulfur-based protecting groups by acyl groups via cap-exchange, special capping agents are used. To retain hydrophobic tag to assist RP HPLC purification, special phosphoramidites are used in the last synthetic cycle. With the sulfur-based groups for protection and linking, oligonucleotide deprotection and cleavage are achieved via oxidation followed by beta-elimination under mild conditions. Therefore, besides for insensitive oligonucleotide synthesis, the embodiments of the invention are capable for the synthesis of oligonucleotide analogs containing sensitive functional groups that cannot survive the harsh conditions used in prior art oligonucleotide synthesis technologies.

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 present invention provides compounds useful as selectively soluble supports (or tags) for oligonucleotide and peptide synthesis that can be removed in a high yield under basic conditions and can afford a resulting product at a high purity in an organic synthesis reaction such as peptide synthesis, oligonucleotide synthesis, and the like. The protecting reagent is a selectively soluble compound of Formula (I) or Formula (II), which is derived from 5-hydroxyisophthalic acid.

The invention discloses synthesis of 3 '-amido-5-nucleotide phosphamide and application of the 3'-amido-5-nucleotide phosphamide in oligonucleotide. The phosphamide modified oligonucleotide comprises at least one of nucleotide structural units shown in a formula I, a formula II, a formula III or a formula IV,..., the formula I, the formula II, the formula III,..., the formula IV. Through specific limitation of a nucleotide phosphamide monomer structure, oligonucleotides are prepared by coupling according to a direction from 5 to 3. Compared with a traditional oligonucleotide molecule, the oligonucleotide molecule provided by the invention contains phosphamide modification and has more excellent effects on pharmaceutical parameters such as enzymatic degradation resistance, stability and the like, in addition, the phosphamide modification can improve the binding performance between the modified oligonucleotide molecule and environmental ions, and the stability of the oligonucleotide molecule is improved. A novel chemical modification strategy is provided for research, development and application of oligonucleotide drugs.

The invention discloses a preparation method and application of a TLR9 targeted CpG oligonucleotide adjuvant, and belongs to the technical field of vaccine adjuvants, and the preparation method comprises the following steps: sequentially adding a phosphoramidite monomer solution, a Unylinker350 carrier, a deprotection reagent, a coupling activator, a sulfo reagent and a capping reagent into an oligonucleotide synthesizer; automatically synthesizing an oligonucleotide fragment in an oligonucleotide synthesizer by adopting a phosphoramidite method to obtain a reaction solution containing the oligonucleotide fragment; carrying out ammonolysis reaction to obtain an ammonolysis crude product; purifying the ammonolysis crude product by using an ion exchange method, collecting components with the purity of more than or equal to 90%, and merging and ultra-filtering; according to the method, the synthesis process conditions are optimized, so that the purity of the finally synthesized CpG oligonucleotide is more than 95%, and the yield is more than 70%.