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    Guide RNAs with Chemical Modification for Prime Editing
    1.
    发明申请
    Guide RNAs with Chemical Modification for Prime Editing 有权

    公开(公告)号：US20230092393A1

    公开(公告)日：2023-03-23

    申请号：US17931535

    申请日：2022-09-12

    Applicant: Agilent Technologies, Inc.

    Inventor： Daniel E. Ryan ,  Robert Kaiser ,  Douglas J. Dellinger

    IPC: C12N15/10 ,  C12N9/22 ,  C12N15/62

    Abstract: Provided herein are compositions and methods for inducing CRISPR/Cas-based editing of a target nucleic acid (e.g., target DNA or target RNA) in vitro or in a cell, using modified prime editing guide RNAs (pegRNAs) that incorporate one or more chemically-modified nucleotides. The modified pegRNAs disclosed herein may be used to induce Cas-mediated incorporation of one or more nucleotide changes and/or targeted mutagenesis of a target nucleic acid. The nucleotide change can include, e.g., one or more nucleotide changes, an insertion of one or more nucleotides, or a deletion of one or more nucleotides.

    Phosphorous protecting groups and methods of preparation and use thereof
    2.
    发明授权
    Phosphorous protecting groups and methods of preparation and use thereof 有权

    公开(公告)号：US11584773B2

    公开(公告)日：2023-02-21

    申请号：US17379288

    申请日：2021-07-19

    Applicant: Agilent Technologies, Inc.

    Inventor： Douglas J. Dellinger ,  Luca Monfregola ,  Marvin Caruthers ,  Mithun Roy

    IPC: C07H19/10 ,  C07H19/23 ,  C07H19/073 ,  C07H19/173 ,  C07H21/04 ,  C40B40/06

    Abstract: Aspects of the present disclosure include compositions that make use of phosphorus and/or nucleobase protecting groups which find use in the synthesis of long polynucleotides. Phosphorus protecting groups are provided that help increase the stepwise coupling yield and/or phosphorous protecting groups that can be removed during the oxidation step. Amidine nucleobase protecting groups are provided that find use in the subject compositions and methods which provides for e.g., increased resistance to depurination during polynucleotide synthesis. In some instances, the methods and compositions disclosed herein utilize a combination of the phosphorus and amidine nucleobase protecting groups in the synthesis of polynucleotides having a sequence of 200 or more monomeric units in length. Also provided are methods for synthesizing a polynucleotide (e.g., a DNA) using one or more compounds disclosed herein.

    Phosphorous protecting groups and methods of preparation and use thereof
    3.
    发明授权
    Phosphorous protecting groups and methods of preparation and use thereof 有权

    公开(公告)号：US11104699B2

    公开(公告)日：2021-08-31

    申请号：US16173143

    申请日：2018-10-29

    Applicant: Agilent Technologies, Inc.

    Inventor： Douglas J. Dellinger ,  Luca Monfregola ,  Marvin Caruthers ,  Mithun Roy

    IPC: C07H19/10 ,  C07H19/23 ,  C07H19/073 ,  C07H19/173 ,  C07H21/04

    Abstract: Aspects of the present disclosure include compositions that make use of phosphorus and/or nucleobase protecting groups which find use in the synthesis of long polynucleotides. Phosphorus protecting groups are provided that help increase the stepwise coupling yield and/or phosphorous protecting groups that can be removed during the oxidation step. Amidine nucleobase protecting groups are provided that find use in the subject compositions and methods which provides for e.g., increased resistance to depurination during polynucleotide synthesis. In some instances, the methods and compositions disclosed herein utilize a combination of the phosphorus and amidine nucleobase protecting groups in the synthesis of polynucleotides having a sequence of 200 or more monomeric units in length. Also provided are methods for synthesizing a polynucleotide (e.g., a DNA) using one or more compounds disclosed herein.

    Protecting groups for “Z nucleotide” and methods thereof
    8.
    发明授权
    Protecting groups for “Z nucleotide” and methods thereof 有权

    公开(公告)号：US09802975B2

    公开(公告)日：2017-10-31

    申请号：US14724613

    申请日：2015-05-28

    Applicant: Agilent Technologies, Inc.

    Inventor： Benjamin D. Lunstad ,  Douglas J. Dellinger

    IPC: C07H5/06 ,  C07H7/06 ,  C07H23/00 ,  C07H21/00 ,  C07D211/00

    CPC classification number: C07H23/00 ,  C07D211/00 ,  C07H7/06 ,  C07H21/00 ,  Y02P20/55

    Abstract: The present disclosure provides nucleoside compounds and oligonucleotides including an unnatural 6-amino-2-pyridone heterocyclic base where the 6-amino and 2-positions are protected. The 2-position of the heterocyclic base can be protected with an acyl-oxy-methyl protecting group. In some embodiments, the protected heterocyclic base has the following structure where AcOM is an acetyl-oxy-methyl group and R is a ribose or deoxyribose sugar: Methods for synthesizing an oligonucleotide are provided in which the subject compounds find use. The method can include protecting an unnatural (e.g., Z) nucleotide with an acetyl-oxy-methyl group; incorporating the protected unnatural nucleotide into a nucleotide sequence on a solid support; and removing the acetyl-oxy-methyl group from the unnatural nucleotide incorporated into the nucleotide sequence. The compounds and methods find use in the synthesis of long oligonucleotides including Z nucleotides.

    PHOSPHOROUS PROTECTING GROUPS AND METHODS OF PREPARATION AND USE THEREOF
    9.
    发明申请
    PHOSPHOROUS PROTECTING GROUPS AND METHODS OF PREPARATION AND USE THEREOF 审中-公开
    磷酸盐保护组及其制备和使用方法

    公开(公告)号：US20150315227A1

    公开(公告)日：2015-11-05

    申请号：US14701288

    申请日：2015-04-30

    Applicant: Agilent Technologies, Inc.

    Inventor： Douglas J. Dellinger ,  Luca Monfregola ,  Marvin Caruthers ,  Mithum Roy

    IPC: C07H19/10 ,  C07H19/23 ,  C07H21/04

    CPC classification number: C07H19/10 ,  C07H19/073 ,  C07H19/173 ,  C07H19/23 ,  C07H21/04 ,  Y02P20/55

    Abstract: Aspects of the present disclosure include compositions that make use of phosphorus and/or nucleobase protecting groups which find use in the synthesis of long polynucleotides. Phosphorus protecting groups are provided that help increase the stepwise coupling yield and/or phosphorous protecting groups that can be removed during the oxidation step. Amidine nucleobase protecting groups are provided that find use in the subject compositions and methods which provides for e.g., increased resistance to depurination during polynucleotide synthesis. In some instances, the methods and compositions disclosed herein utilize a combination of the phosphorus and amidine nucleobase protecting groups in the synthesis of polynucleotides having a sequence of 200 or more monomeric units in length. Also provided are methods for synthesizing a polynucleotide (e.g., a DNA) using one or more compounds disclosed herein.

    Abstract translation: 本公开的方面包括利用可用于合成长多核苷酸的磷和/或核碱基保护基的组合物。 提供了磷保护基团，其有助于增加在氧化步骤期间可以除去的逐步偶联产率和/或磷保护基团。 提供了在本发明组合物和方法中使用的脒核碱基保护基，其提供例如在多核苷酸合成期间提高的去除抗性。 在一些情况下，本文公开的方法和组合物在合成具有200个或更多个单体单元长度的序列的多核苷酸中的磷和脒核碱基保护基团的组合。 还提供了使用本文公开的一种或多种化合物合成多核苷酸（例如DNA）的方法。

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