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    Applications of Streptococcus-derived Cas9 nucleases on minimal Adenine-rich PAM targets
    1.
    发明申请
    Applications of Streptococcus-derived Cas9 nucleases on minimal Adenine-rich PAM targets 有权

    公开(公告)号：US20250154513A1

    公开(公告)日：2025-05-15

    申请号：US18927790

    申请日：2024-10-25

    Applicant: Massachusetts Institute of Technology

    Inventor： Pranam Chatterjee ,  Noah Michael Jakimo ,  Joseph M. Jacobson

    IPC: C12N15/74 ,  C12N9/22 ,  C12N15/11

    Abstract: Applications of a Streptococcus Cas9 ortholog from Streptococcus macacae (Smac Cas9), possessing minimal adenine-rich PAM specificity, include an isolated Streptococcus macacae Cas9 protein or transgene expression thereof, a CRISPR-associated DNA endonuclease with PAM interacting domain amino acid sequences that are at least 80% identical to that of the isolated Streptococcus macacae Cas9 protein, and an isolated, engineered Streptococcus pyogenes Cas9 (Spy Cas9) protein with a PID as either the PID amino acid composition of the isolated Streptococcus macacae Cas9 (Smac Cas9) protein or of a CRISPR-associated DNA endonuclease with PID amino acid sequences that are at least 80% identical to that of the isolated Streptococcus macacae Cas9 protein. A method for altering expression of at least one gene product employs Streptococcus macacae Cas9 endonucleases in complex with guide RNA, for specific recognition and activity on a DNA target immediately upstream of either an “NAA” or “NA” or “NAAN” PAM sequence.

    Applications of engineered Streptococcus canis Cas9 variants on single-base PAM targets
    2.
    发明授权
    Applications of engineered Streptococcus canis Cas9 variants on single-base PAM targets 有权

    公开(公告)号：US11453865B2

    公开(公告)日：2022-09-27

    申请号：US16689071

    申请日：2019-11-19

    Applicant: Massachusetts Institute of Technology

    Inventor： Pranam Chatterjee ,  Noah Michael Jakimo ,  Joseph M. Jacobson

    IPC: C12N9/22 ,  C12N15/11 ,  C12N15/85

    Abstract: Engineered Streptococcus canis Cas9 (ScCas9) variants include an ScCas9 protein with its PID being the PID amino acid composition of Streptococcus pyogenes Cas9 (SpCas9)-NG, an ScCas9 protein having a threonine-to-lysine substitution mutation at position 1227 in its amino acid sequence (Sc+), and an ScCas9 protein having a threonine-to-lysine substitution mutation at position 1227 and a substitution of residues ADKKLRKRSGKLATE [SEQ ID No. 4] in position 365-379 in the ScCas9 open reading frame (Sc++). Also included are CRISPR-associated DNA endonucleases with a PAM specificity of 5′-NG-3′ or 5′-NNG-3′ and a method of altering expression of a gene product by utilizing the engineered ScCas9 variants.

    Applications of Streptococcus-derived Cas9 nucleases on minimal Adenine-rich PAM targets
    3.
    发明申请
    Applications of Streptococcus-derived Cas9 nucleases on minimal Adenine-rich PAM targets 有权

    公开(公告)号：US20220162620A1

    公开(公告)日：2022-05-26

    申请号：US17053713

    申请日：2019-05-06

    Applicant: Massachusetts Institute of Technology

    Inventor： Pranam Chatterjee ,  Noah Michael Jakimo ,  Joseph M Jacobson

    IPC: C12N15/74 ,  C12N9/22 ,  C12N15/11

    Abstract: Applications of a Streptococcus Cas9 ortholog from Streptococcus macacae (Smac Cas9), possessing minimal adenine-rich PAM specificity, include an isolated Streptococcus macacae Cas9 protein or transgene expression thereof, a CRISPR-associated DNA endonuclease with PAM interacting domain amino acid sequences that are at least 80% identical to that of the isolated Streptococcus macacae Cas9 protein, and an isolated, engineered Streptococcus pyogenes Cas9 (Spy Cas9) protein with a PID as either the PID amino acid composition of the isolated Streptococcus macacae Cas9 (Smac Cas9) protein or of a CRISPR-associated DNA endonuclease with PID amino acid sequences that are at least 80% identical to that of the isolated Streptococcus macacae Cas9 protein. A method for altering expression of at least one gene product employs Streptococcus macacae Cas9 endonucleases in complex with guide RNA, for specific recognition and activity on a DNA target immediately upstream of either an “NAA” or “NA” or “NAAN” PAM sequence.

    Applications of engineered
    7.
    发明授权
    Applications of engineered 有权

    公开(公告)号：US11697808B2

    公开(公告)日：2023-07-11

    申请号：US17855507

    申请日：2022-06-30

    Applicant: Massachusetts Institute of Technology

    Inventor： Pranam Chatterjee ,  Noah Michael Jakimo ,  Joseph M. Jacobson

    IPC: C12N9/22 ,  C12N15/11 ,  C12N15/85

    CPC classification number: C12N9/22 ,  C12N15/11 ,  C12N15/85 ,  C12N2310/20 ,  C12N2800/80

    Abstract: Engineered Streptococcus canis Cas9 (ScCas9) variants include an ScCas9 protein with its PID being the PID amino acid composition of Streptococcus pyogenes Cas9 (SpCas9)-NG, an ScCas9 protein having a threonine-to-lysine substitution mutation at position 1227 in its amino acid sequence (Sc+), and an ScCas9 protein having a threonine-to-lysine substitution mutation at position 1227 and a substitution of residues ADKKLRKRSGKLATE [SEQ ID No. 4] in position 365-379 in the ScCas9 open reading frame (Sc++). Also included are CRISPR-associated DNA endonucleases with a PAM specificity of 5′-NG-3′ or 5′-NNG-3′ and a method of altering expression of a gene product by utilizing the engineered ScCas9 variants.

    Applications of Engineered Streptococcus Canis Cas9 Variants on Single-Base PAM Targets
    8.
    发明申请
    Applications of Engineered Streptococcus Canis Cas9 Variants on Single-Base PAM Targets 有权

    公开(公告)号：US20230067345A1

    公开(公告)日：2023-03-02

    申请号：US17855507

    申请日：2022-06-30

    Applicant: Massachusetts Institute of Technology

    Inventor： Pranam Chatterjee ,  Noah Michael Jakimo ,  Joseph M. Jacobson

    IPC: C12N9/22 ,  C12N15/11 ,  C12N15/85

    Abstract: Engineered Streptococcus canis Cas9 (ScCas9) variants include an ScCas9 protein with its PID being the PID amino acid composition of Streptococcus pyogenes Cas9 (SpCas9)-NG, an ScCas9 protein having a threonine-to-lysine substitution mutation at position 1227 in its amino acid sequence (Sc+), and an ScCas9 protein having a threonine-to-lysine substitution mutation at position 1227 and a substitution of residues ADKKLRKRSGKLATE [SEQ ID No. 4] in position 365-379 in the ScCas9 open reading frame (Sc++). Also included are CRISPR-associated DNA endonucleases with a PAM specificity of 5′-NG-3′ or 5′-NNG-3′ and a method of altering expression of a gene product by utilizing the engineered ScCas9 variants.

    DIRECTED ENDONUCLEASES FOR REPEATABLE NUCLEIC ACID CLEAVAGE
    9.
    发明申请
    DIRECTED ENDONUCLEASES FOR REPEATABLE NUCLEIC ACID CLEAVAGE 审中-公开
    用于可重复核酸清除的指导性内切剂

    公开(公告)号：US20160017393A1

    公开(公告)日：2016-01-21

    申请号：US14800096

    申请日：2015-07-15

    Applicant: Massachusetts Institute of Technology

    Inventor： Joseph M. Jacobson ,  Noah Michael Jakimo

    IPC: C12P19/34

    CPC classification number: C12P19/34 ,  C07K2319/00 ,  C07K2319/81 ,  C12N9/22 ,  C12N2310/20

    Abstract: The invention provides compositions and methods for repeatable directed endonucleases (RDEs) and methods for repeatedly, and specifically cleaving DNA offset from the RDE's DNA recognition sequence on the target nucleic acid rather than within the DNA recognition sequence. Conservation of the recognition sequence of the target nucleic acid enables for re-localization of an RDE back to the DNA recognition sequence for further cleavage. The RDEs and methods of the invention are useful in applications including, but not limited to, recording data into a genome, timing the order of biochemical pathway events, efficient genome engineering and encoding lagged cellular death.

    Abstract translation: 本发明提供了用于重复定向内切核酸酶（RDEs）的组合物和方法以及用于重复且特异性地切割来自目标核酸而不是在DNA识别序列内的RDE的DNA识别序列的DNA偏移的方法。 保护靶核酸的识别序列能够使RDE重新定位回到DNA识别序列以进一步切割。 本发明的RDEs和方法可用于包括但不限于将数据记录到基因组中，定时生化通路事件的顺序，有效的基因组工程和编码滞后的细胞死亡的应用。

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