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公开(公告)号：US11492667B2
公开(公告)日：2022-11-08
申请号：US17342216
申请日：2021-06-08
Applicant: New England Biolabs, Inc.
Inventor： Sriharsa Pradhan , Hang Gyeong Chin , George R. Feehery , Shuang-Yong Xu , Vishnu Udayakumaran Nair Sunitha Kumary , Julie Beaulieu , Pierre O. Esteve
IPC: C12Q1/6869 , C07K14/31 , C12N9/22
Abstract: Compositions, methods and kits are provided for identifying the presence and location of a target in chromosomal DNA. A nicking endonuclease fused to a binding domain that binds to a constant region of an antibody (NEFP) is provided that may be used for binding to a target directly or via an antibody that binds to the target. The target may be a protein or structural feature of the DNA and its presence and location may correspond to a phenotype and/or pathology in a biopsy or other cell sample for diagnostic purposes. The background is reduced by the addition of a glycoaminoglycan (GAG) that reversibly inhibits binding of the NEFP to DNA. Nick translation in the presence of a strand displacing polymerase enables the incorporation of tagged nucleotides that (i) blocks re-nicking; (ii) facilitates immobilization of DNA fragments around the target for sequencing; and/or (iii) enables dye labelling of the chromosomal DNA within the cell nuclei for analysis by microscopy.
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公开(公告)号：US09856464B2
公开(公告)日：2018-01-02
申请号：US15133093
申请日：2016-04-19
Applicant: New England Biolabs, Inc.
Inventor： Zhenyu Zhu , Aine Quimby , Shengxi Guan , Dapeng Sun , Yishu Huang , Xuhui Lai , Siu-hong Chan , Xianghui Li , Shuang-Yong Xu , Chunhua Zhang
CPC classification number: C12N9/22 , C12N9/16 , C12Q1/34 , C12Q1/44 , C12Q1/68 , C12Y301/21004 , G01N2333/916 , G01N2333/922
Abstract: Methods and compositions are provided for engineering mutant enzymes with reduced star activity where the mutant enzymes have a fidelity index (FI) in a specified buffer that is greater than the FI of the non-mutated enzyme in the same buffer.
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公开(公告)号：US20160289660A1
公开(公告)日：2016-10-06
申请号：US15133093
申请日：2016-04-19
Applicant: New England Biolabs, Inc.
Inventor： Zhenyu Zhu , Aine Quimby , Shengxi Guan , Dapeng Sun , Yishu Huang , Xuhui Lai , Siu-hong Chan , Xianghui Li , Shuang-Yong Xu , Chunhua Zhang
CPC classification number: C12N9/22 , C12N9/16 , C12Q1/34 , C12Q1/44 , C12Q1/68 , C12Y301/21004 , G01N2333/916 , G01N2333/922
Abstract: Methods and compositions are provided for engineering mutant enzymes with reduced star activity where the mutant enzymes have a fidelity index (FI) in a specified buffer that is greater than the FI of the non-mutated enzyme in the same buffer.
Abstract translation: 提供了用于工程化具有降低的星形活性的突变酶的方法和组合物，其中突变酶在特定缓冲液中具有比同一缓冲液中非突变酶的FI高的保真度指数（FI）。
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公开(公告)号：US09074196B2
公开(公告)日：2015-07-07
申请号：US13736529
申请日：2013-01-08
Applicant: New England Biolabs, Inc.
Inventor： Zhenyu Zhu , Aine Quimby , Shuang-Yong Xu , Shengxi Guan , Hua Wei , Penghua Zhang , Dapeng Sun , Siu-hong Chan
CPC classification number: C12N15/1058 , C12N9/16 , C12N9/22 , C12N15/1034 , C12N15/1086
Abstract: Compositions and methods are provided for enzymes with altered properties that involve a systematic approach to mutagenesis and a screening assay that permits selection of the desired proteins. Embodiments of the method are particularly suited for modifying specific properties of restriction endonucleases such as star activity. The compositions includes restriction endonucleases with reduced star activity as defined by an overall fidelity index improvement factor.
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公开(公告)号：US20140154779A1
公开(公告)日：2014-06-05
申请号：US14137636
申请日：2013-12-20
Applicant: NEW ENGLAND BIOLABS, INC.
Inventor： Zhenyu Zhu , Aine Quimby , Shengxi Guan , Dapeng Sun , Yishu Huang , Xuhui Lai , Siu-hong Chan , Xianghui Li , Shuang-Yong Xu , Chunhua Zhang
IPC: C12N9/16
CPC classification number: C12N9/22 , C12N9/16 , C12Q1/34 , C12Q1/44 , C12Q1/68 , C12Y301/21004 , G01N2333/916 , G01N2333/922
Abstract: Methods and compositions are provided for engineering mutant enzymes with reduced star activity where the mutant enzymes have a fidelity index (FI) in a specified buffer that is greater than the FI of the non-mutated enzyme in the same buffer.
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Patent Agency Ranking
公开(公告)号：US08628945B2
公开(公告)日：2014-01-14
申请号：US13736170
申请日：2013-01-08
Applicant: New England Biolabs, Inc.
Inventor： Zhenyu Zhu , Aine Quimby , Shuang-Yong Xu , Shengxi Guan , Hua Wei , Penghua Zhang , Dapeng Sun , Siu-hong Chan
IPC: C12N9/22
CPC classification number: C12N15/1058 , C12N9/16 , C12N9/22 , C12N15/1034 , C12N15/1086
Abstract: Compositions and methods are provided for enzymes with altered properties that involve a systematic approach to mutagenesis and a screening assay that permits selection of the desired proteins. Embodiments of the method are particularly suited for modifying specific properties of restriction endonucleases such as star activity. The compositions includes restriction endonucleases with reduced star activity as defined by an overall fidelity index improvement factor.
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公开(公告)号：US20130115680A1
公开(公告)日：2013-05-09
申请号：US13736170
申请日：2013-01-08
Applicant: NEW ENGLAND BIOLABS, INC.
Inventor： Zhenyu Zhu , Aine Quimby , Shuang-Yong Xu , Shengxi Guan , Hua Wei , Penghua Zhang , Dapeng Sun , Siu-hong Chan
IPC: C12N9/22
CPC classification number: C12N15/1058 , C12N9/16 , C12N9/22 , C12N15/1034 , C12N15/1086
Abstract: Compositions and methods are provided for enzymes with altered properties that involve a systematic approach to mutagenesis and a screening assay that permits selection of the desired proteins. Embodiments of the method are particularly suited for modifying specific properties of restriction endonucleases such as star activity. The compositions includes restriction endonucleases with reduced star activity as defined by an overall fidelity index improvement factor.
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公开(公告)号：US20240352447A1
公开(公告)日：2024-10-24
申请号：US18750621
申请日：2024-06-21
Applicant: New England Biolabs, Inc.
Inventor： Zhenyu Zhu , Aine Quimby , Shuang-Yong Xu , Shengxi Guan , Hua Wei , Penghua Zhang , Dapeng Sun , Siu-hong Chan
CPC classification number: C12N15/1058 , C12N9/16 , C12N9/22 , C12N15/1034 , C12N15/1086
Abstract: Compositions and methods are provided for enzymes with altered properties that involve a systematic approach to mutagenesis and a screening assay that permits selection of the desired proteins. Embodiments of the method are particularly suited for modifying specific properties of restriction endonucleases such as star activity. The compositions includes restriction endonucleases with reduced star activity as defined by an overall fidelity index improvement factor.
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公开(公告)号：US20240352437A1
公开(公告)日：2024-10-24
申请号：US18762074
申请日：2024-07-02
Applicant: New England Biolabs, Inc.
Inventor： Zhenyu Zhu , Aine Quimby , Shengxi Guan , Dapeng Sun , Yishu Huang , Xuhui Lai , Siu-hong Chan , Xianghui Li , Shuang-Yong Xu , Chunhua Zhang
CPC classification number: C12N9/22 , C12N9/16 , C12Q1/34 , C12Q1/44 , C12Q1/68 , C12Y301/21004 , G01N2333/916 , G01N2333/922
Abstract: Methods and compositions are provided for engineering mutant enzymes with reduced star activity where the mutant enzymes have a fidelity index (FI) in a specified buffer that is greater than the FI of the non-mutated enzyme in the same buffer.
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公开(公告)号：US20230123635A1
公开(公告)日：2023-04-20
申请号：US17907482
申请日：2021-06-08
Applicant: New England Biolabs, Inc.
Inventor： Sriharsa Pradhan , Hang Gyeong Chin , George R. Feehery , Shuang-Yong Xu , Vishnu Udayakumaran Nair Sunitha Kumary , Julie Beaulieu
IPC: C12Q1/6841
Abstract: Compositions, methods and kits are provided for identifying the presence and location of a target in chromosomal DNA. A nicking endonuclease fused to a binding domain that binds to a constant region of an antibody (NEFP) is provided that may be used for binding to a target directly or via an antibody that binds to the target. The target may be a protein or structural feature of the DNA and its presence and location may correspond to a phenotype and/or pathology in a biopsy or other cell sample for diagnostic purposes. The background is reduced by the addition of a glycoaminoglycan (GAG) that reversibly inhibits binding of the NEFP to DNA. Nick translation in the presence of a strand displacing polymerase enables the incorporation of tagged nucleotides that (i) blocks re-nicking; (ii) facilitates immobilization of DNA fragments around the target for sequencing; and/or (iii) enables dye labelling of the chromosomal DNA within the cell nuclei for analysis by microscopy.
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