公开(公告)号：US10946108B2

公开(公告)日：2021-03-16

申请号：US14970967

申请日：2015-12-16

申请人： The Broad Institute, Inc. ,  Massachusetts Institute of Technology ,  President and Fellows of Harvard College

发明人： Feng Zhang ,  Le Cong ,  Fei Ran ,  Matthias Heidenreich ,  Lukasz Swiech

IPC分类号： A61K48/00 ,  C12N9/22 ,  C12N15/63 ,  C12N15/10 ,  C12N7/00 ,  C12N15/86

摘要： The invention provides for delivery, engineering and optimization of systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are delivery systems and tissues or organ which are targeted as sites for delivery. Also provided are vectors and vector systems some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells to ensure enhanced specificity for target recognition and avoidance of toxicity and to edit or modify a target site in a genomic locus of interest to alter or improve the status of a disease or a condition.

公开(公告)号：US20190218276A1

公开(公告)日：2019-07-18

申请号：US16087036

申请日：2016-10-27

申请人： THE BROAD INSTITUTE, INC. ,  MASSACHUSETTS INSTITUTE OF TECHNOLOGY ,  PRESIDENT AND FELLOWS OF HARVARD COLLEGE

发明人： Aviv Regev ,  Feng Zhang ,  Naomi Habib ,  Yinqing Li ,  Matthias Heidenreich ,  Lukasz Swiech ,  Anindita Basu ,  David Weitz ,  Inbal Avraham Davidi

IPC分类号： C07K16/10 ,  C12N15/10 ,  C12Q1/686 ,  C12Q1/6869

CPC分类号： C07K16/1036 ,  C12N15/1093 ,  C12N15/1096 ,  C12Q1/686 ,  C12Q1/6869 ,  C12Q2537/165 ,  C12Q2563/179 ,  C12Q2565/514 ,  C12Q2565/629

摘要： Transcriptomes of individual neurons provide rich information about cell types and dynamic states. However, it is difficult to capture rare dynamic processes, such as adult neurogenesis, because isolation from dense adult tissue is challenging, and markers for each phase are limited. Here, Applicants developed Nuc-seq, Div-Seq, and Dronc-Seq. Div-seq combines Nuc-Seq, a scalable single nucleus RNA-Seq method, with EdU-mediated labeling of proliferating cells. Nuc-Seq can sensitively identify closely related cell types within the adult hippocampus. Div-Seq can track transcriptional dynamics of newborn neurons in an adult neurogenic region in the hippocampus. Dronc-Seq uses a microfluidic device to co-encapsulate individual nuclei in reverse emulsion aqueous droplets in an oil medium together with one uniquely barcoded mRNA-capture bead. Finally, Applicants found rare adult newborn GABAergic neurons in the spinal cord, a non-canonical neurogenic region. Taken together, Nuc-Seq, Div-Seq and Dronc-Seq allow for unbiased analysis of any complex tissue.

公开(公告)号：US12060412B2

公开(公告)日：2024-08-13

申请号：US16087036

申请日：2016-10-27

申请人： THE BROAD INSTITUTE, INC. ,  MASSACHUSETTS INSTITUTE OF TECHNOLOGY ,  PRESIDENT AND FELLOWS OF HARVARD COLLEGE

发明人： Aviv Regev ,  Feng Zhang ,  Naomi Habib ,  Yinqing Li ,  Matthias Heidenreich ,  Lukasz Swiech ,  Anindita Basu ,  David Weitz ,  Inbal Avraham Davidi

IPC分类号： C07K16/10 ,  C12N15/10 ,  C12Q1/686 ,  C12Q1/6869

CPC分类号： C07K16/1036 ,  C12N15/1093 ,  C12N15/1096 ,  C12Q1/686 ,  C12Q1/6869 ,  C12Q2537/165 ,  C12Q2563/179 ,  C12Q2565/514 ,  C12Q2565/629 ,  C12Q1/6869 ,  C12Q2537/165 ,  C12Q2563/179 ,  C12Q2565/514 ,  C12Q2565/629

摘要： Transcriptomes of individual neurons provide rich information about cell types and dynamic states. However, it is difficult to capture rare dynamic processes, such as adult neurogenesis, because isolation from dense adult tissue is challenging, and markers for each phase are limited. Here, Applicants developed Nuc-seq, Div-Seq, and Dronc-Seq. Div-seq combines Nuc-Seq, a scalable single nucleus RNA-Seq method, with EdU-mediated labeling of proliferating cells. Nuc-Seq can sensitively identify closely related cell types within the adult hippocampus. Div-Seq can track transcriptional dynamics of newborn neurons in an adult neurogenic region in the hippocampus. Dronc-Seq uses a microfluidic device to co-encapsulate individual nuclei in reverse emulsion aqueous droplets in an oil medium together with one uniquely barcoded mRNA-capture bead. Finally, Applicants found rare adult newborn GABAergic neurons in the spinal cord, a non-canonical neurogenic region. Taken together, Nuc-Seq, Div-Seq and Dronc-Seq allow for unbiased analysis of any complex tissue.

公开(公告)号：US20210361779A1

公开(公告)日：2021-11-25

申请号：US17201347

申请日：2021-03-15

申请人： The Broad Institute, Inc. ,  Massachusetts Institute of Technology ,  President and Fellows of Harvard College

发明人： Feng Zhang ,  Le Cong ,  Fei Ran ,  Matthias Heidenreich ,  Lukasz Swiech

IPC分类号： A61K48/00 ,  C12N9/22 ,  C12N15/63 ,  C12N15/10 ,  C12N7/00 ,  C12N15/86

摘要： The invention provides for delivery, engineering and optimization of systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are delivery systems and tissues or organ which are targeted as sites for delivery. Also provided are vectors and vector systems some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells to ensure enhanced specificity for target recognition and avoidance of toxicity and to edit or modify a target site in a genomic locus of interest to alter or improve the status of a disease or a condition.

公开(公告)号：US20160340661A1

公开(公告)日：2016-11-24

申请号：US15179799

申请日：2016-06-10

申请人： The Broad Institute Inc. ,  Massachusetts Institute of Technology

发明人： Le Cong ,  David Benjamin Turitz Cox ,  Matthias Heidenreich ,  Randall Jeffrey Platt ,  Lukasz Swiech ,  Feng Zhang

IPC分类号： C12N9/22 ,  A61K9/00 ,  A61K38/46

摘要： The invention provides for delivery, engineering and optimization of systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are delivery systems and tissues or organ which are targeted as sites for delivery. Also provided are vectors and vector systems some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells to ensure enhanced specificity for target recognition and avoidance of toxicity and to edit or modify a target site in a genomic locus of interest to alter or improve the status of a disease or a condition.

摘要翻译： 本发明提供了用于操纵靶序列的序列和/或活性的系统，方法和组合物的递送，工程和优化。 提供作为递送站点的递送系统和组织或器官。 还提供了载体和载体系统，其中一些编码CRISPR复合物的一个或多个组分，以及用于设计和使用这些载体的方法。 还提供了在真核细胞中引导CRISPR复合物形成的方法，以确保增强的靶标识别和避免毒性的特异性，以及编辑或修饰感兴趣的基因组基因座中的靶位点以改变或改善疾病或病症的状态。

公开(公告)号：US11407985B2

公开(公告)日：2022-08-09

申请号：US15179799

申请日：2016-06-10

申请人： The Broad Institute, Inc. ,  Massachusetts Institute of Technology

发明人： Le Cong ,  David Benjamin Turitz Cox ,  Matthias Heidenreich ,  Randall Jeffrey Platt ,  Lukasz Swiech ,  Feng Zhang

IPC分类号： A61K48/00 ,  C12N9/22 ,  C12N15/90 ,  C12N15/10 ,  A61K9/00 ,  A61K38/46

摘要： The invention provides for delivery, engineering and optimization of systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are delivery systems and tissues or organ which are targeted as sites for delivery. Also provided are vectors and vector systems some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells to ensure enhanced specificity for target recognition and avoidance of toxicity and to edit or modify a target site in a genomic locus of interest to alter or improve the status of a disease or a condition.