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    Nanoparticle chains and Preparation Thereof
    2.
    发明申请
    Nanoparticle chains and Preparation Thereof 审中-公开

    公开(公告)号：US20170190802A1

    公开(公告)日：2017-07-06

    申请号：US15410756

    申请日：2017-01-19

    Applicant: Massachusetts Institute of Technology

    Inventor： Joseph M. Jacobson ,  David W. Mosley ,  Kie-Moon Sung

    IPC: C08B37/16

    CPC classification number: C08B37/0012 ,  B05D1/00 ,  C01B13/00 ,  C08B37/0015 ,  Y10S977/778 ,  Y10S977/882 ,  Y10T428/25 ,  Y10T428/2991

    Abstract: Fabrication and arrangement of nanoparticles into one-dimensional linear chains is achieved by successive chemical reactions, each reaction adding one or more nanoparticles by building onto exposed, unprotected linker functionalities. Optionally, protecting groups may be used to control and organize growth. Nanoparticle spheres are functionalized in a controlled manner in order to enable covalent linkages. Functionalization of nanoparticles is accomplished by either ligand exchange or chemical modification of the terminal functional groups of the capping ligand. Nanoparticle chains are obtained by a variety of connectivity modes such as direct coupling, use of linker molecules, and use of linear polymeric templates. In particular, a versatile building block system is obtained through controlled monofunctionalization of nanoparticles.

    DIRECTED ENDONUCLEASES FOR REPEATABLE NUCLEIC ACID CLEAVAGE
    3.
    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。

    Minimal Peptide Fusions for Targeted Intracellular Degradation of FOXP3
    6.
    发明公开
    Minimal Peptide Fusions for Targeted Intracellular Degradation of FOXP3 审中-公开

    公开(公告)号：US20230257725A1

    公开(公告)日：2023-08-17

    申请号：US17972132

    申请日：2022-10-24

    Applicant: Massachusetts Institute of Technology

    Inventor： Pranam Chatterjee ,  Joseph M. Jacobson

    IPC: C12N9/48

    CPC classification number: C12N9/48 ,  C12Y304/17023

    Abstract: Peptide-E3 ubiquitin ligase fusions representing minimal protein to proteasome linkers are specifically targeted to degrade endogenous FOXP3 proteins in regulatory T cells. An engineered peptide for functional inactivation of a target regulatory T cell includes a fusion protein comprising a targeting domain and a ubiquitin ligase recruiting domain, wherein the targeting domain is engineered to bind FOXP3 of the target regulatory T cell for mediated degradation by the ubiquitin-proteosome pathway. The targeting domain may comprise a peptide having amino acid [SEQ ID No. 3], [SEQ ID No. 4], [SEQ ID No. 5], [SEQ ID No. 6], or [SEQ ID No. 7]. The ubiquitin ligase recruiting domain recruits an E3 ubiquitin ligase, which may be CHIPΔTPR [SEQ ID No. 2]. An engineered minimal, specific, nucleotide-encodable, FOXP3 protein to proteasome linker comprises a peptide-E3 ubiquitin ligase fusion in which the peptide binds to FOXP3. A method for treatment includes administering to a subject an engineered peptide-based therapeutic or pharmaceutically acceptable salt thereof, wherein the engineered peptide-based therapeutic comprises a peptide fusion of a targeting domain and a ubiquitin ligase recruiting domain, and wherein the targeting domain is engineered to bind FOXP3 of at least one regulatory T cell for mediated degradation by the ubiquitin-proteosome pathway.

    Methods for High Fidelity Production of Long Nucleic Acid Molecules
    8.
    发明申请
    Methods for High Fidelity Production of Long Nucleic Acid Molecules 审中-公开

    公开(公告)号：US20170137858A1

    公开(公告)日：2017-05-18

    申请号：US15359560

    申请日：2016-11-22

    Applicant: Massachusetts Institute of Technology

    Inventor： Peter A. Carr ,  Brian Y. Chow ,  Joseph M. Jacobson ,  David W. Mosley ,  Christopher Emig

    IPC: C12P19/34 ,  C12Q1/68

    CPC classification number: C12P19/34 ,  C12Q1/6825 ,  C12Q1/6834 ,  C12Q1/6848 ,  C12Q2565/518 ,  C12Q2565/513 ,  C12Q2521/514

    Abstract: In a method for generating an elongated nucleic acid molecule, a nucleic acid addition of a first nucleic acid molecule attached to a first 3′ or 5′ protecting group to a nucleic acid immobilized on a surface produces an intermediate-length immobilized nucleic acid. The first protecting group is dissociated from the first nucleic acid molecule. A second nucleic acid molecule that is attached to a second associated a 3′ or 5′ associated protecting group is added to the intermediate-length nucleic acid. The second associated protecting group is dissociated from the second nucleic acid molecule. A sequentially-extended elongated immobilized nucleic acid molecule having a desired sequence and length is produced by sequentially extending the intermediate-length immobilized nucleic acid by adding additional nucleic acid molecules with associated protecting groups to the intermediate-length nucleic acid and dissociating the associated protecting group after each addition.

    Applications of Streptococcus-derived Cas9 nucleases on minimal Adenine-rich PAM targets
    9.
    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
    10.
    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.

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