57 results about "Oligonucleotide synthesis" patented technology

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 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.

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 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.

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.

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.

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.

Provided is a highly fat-soluble nucleotide block for the synthesis of an oligonucleotide having a structure represented by formula (III) or (VI). In formulae (III) and (VI), R1 is at least any one of Tr, MMTr, DMTr, TMTr, TBDPS, a benzoyl group, an isobutyryl group, dmf, and Ac, and in formulae (III) and (VI), when OR2 and Y are both OMMTr, ODMTr, and OTMTr, R1 may not have a protecting group; and R2 is any one of Tr, MMTr, DMTr, TMTr, TBDPS, Fmoc, and 1-pyrenyl methyl.

This invention provides an adaptive dynamic reservoir, a reservoir system, and an oligonucleotide synthesis system thereof, including an adaptive dynamic reservoir, a reservoir system based on the adaptive dynamic reservoir, and an oligonucleotide synthesis system incorporating the adaptive dynamic reservoir. This invention has the advantages of being able to temporarily store monomers and activators exceeding column volume during the coupling step and participate in the cyclic step; and being able to drain them before the washing step for easy cleaning.

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.

Functionalized solid supports are useful in the synthesis of oligonucleotides. The functionalized solid supports contain an extended linker to a terminal functional group or a first nucleotide or nucleoside moiety. The extended linker permits oligonucleotide synthesis to take place at a greater distance from the solid support with greater efficiency.

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.

To provide an efficient solid phase oligonucleotide synthesis (SPOS) method.SOLUTION: A method of adding an oligonucleotide to a solid support in a bed reactor, the method comprising: removing a protecting group from a 5' - position of an oligonucleotide bound to the solid support; and adding an activated amidite solution to the bed reactor, wherein: Wherein the activated amidite solution comprises an amidite and flows up and down or is fluidized by nitrogen bubbling or other agitation in the bed reactor and reacts at the 5 ' position of the oligonucleotide, wherein the phosphite linkage found in the amidite comprises a P atom in oxidation state III, and then converting the P atom from oxidation state III to oxidation state V.SELECTED DRAWING: None

This continuous oligonucleotide synthesis device is provided with: a raw material container for accommodating a protective nucleoside block as a raw material; a first reagent container containing a reagent for protecting the chain extension of the nucleoside block; a second reagent container for accommodating a reagent for a reaction of oxidizing or vulcanizing a phosphoric acid bond of a nucleotide chain extender obtained by chain extension; and a third reagent container containing a reagent for a reaction for deprotecting the oxidized or vulcanized nucleotide chain extender, and synthesizing oligonucleotides in a flow format by continuously supplying the raw material, the first reagent, and the second reagent.

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 a step of capping reaction solution recycling. In some embodiments, recycling the capping reaction solution requires a significantly reduced amount of capping solution during the entire SPOS process relative to SPOS processes where the capping step of each cycle is carried out using fresh oxidation solution. In some embodiments, oligonucleotides produced using systems and methods of the disclosure have higher purity than oligonucleotides produced using conventional SPOS techniques