公开(公告)号：US11766488B2

公开(公告)日：2023-09-26

申请号：US16315458

申请日：2017-07-05

申请人： The Johns Hopkins University

发明人： Vinod Jaskula-Ranga ,  Donald Zack ,  Derek Welsbie

IPC分类号： A61K48/00 ,  A61K9/00 ,  C12N15/86

CPC分类号： A61K48/005 ,  A61K9/0014 ,  A61K9/0019 ,  C12N15/86 ,  C12N2310/20 ,  C12N2330/51 ,  C12N2710/10344 ,  C12N2750/14143

摘要： The presently disclosed subject matter provides compositions and methods comprising improvements of a CRISPR system (e.g. CRISPR associated (Cas) 9 (CRISPR-Cas9, non-Cas9 CRISPR systems). Such compositions may comprise modifications to the H1 promoter region, addition of 5′UTR modifications, different orthologous sequences of the H1 promoter, novel compact bidirectional promoter sequences with both pol II and pol III activity, addition of Kozak consensus sequences, termination sequences, addition of conditional pol II/pol III bidirectional promoter expression, addition of a donor template sequence for correcting mutations, or combinations thereof. Other aspects of the invention relate to modifications to Cas9 through post-transcriptional cell-cycle regulation fusions, engineered partial target sites such that the nuclease can bind without DNA cleavage, auto-regulation sites, and N-terminal modifications to modulate half-life.

公开(公告)号：US11850289B2

公开(公告)日：2023-12-26

申请号：US16851636

申请日：2020-04-17

申请人： The Johns Hopkins University

发明人： Vinod Jaskula-Ranga ,  Donald Zack

IPC分类号： A61K48/00 ,  C12N15/85 ,  C12N15/113 ,  C12N15/63 ,  C12N15/90 ,  C12N15/11 ,  C12N15/861 ,  C12N15/86 ,  C12Q1/68

CPC分类号： A61K48/0066 ,  C12N15/111 ,  C12N15/1138 ,  C12N15/63 ,  C12N15/85 ,  C12N15/86 ,  C12N15/8616 ,  C12N15/907 ,  C07K2319/10 ,  C07K2319/60 ,  C12N2310/10 ,  C12N2310/20 ,  C12N2330/51 ,  C12N2750/14133 ,  C12N2750/14143 ,  C12N2830/205 ,  C12N2830/85

摘要： The presently disclosed subject matter provides compositions and methods for the expression of CRISPR guide RNAs using the H1 promoter. In particular, compositions and methods are provided for the use of the H1 promoter to express CRISPR guide RNA (gRNA) with altered specificity of the 5′ nucleotide, as well as use of the H1 promoter sequence as a bidirectional promoter to express Cas9 nuclease and the gRNA simultaneously. Compositions and methods are also provided for the expression and regulation of gRNA expression in vivo through the use of RNA ribozymes and regulatable aptazymes.

公开(公告)号：US10668173B2

公开(公告)日：2020-06-02

申请号：US16445352

申请日：2019-06-19

申请人： The Johns Hopkins University

发明人： Vinod Jaskula-Ranga ,  Donald Zack

IPC分类号： A61K48/00 ,  C12N15/86 ,  C12N15/113 ,  C12N15/63 ,  C12N15/85 ,  C12N15/90 ,  C12N15/11 ,  C12N15/861

摘要： The presently disclosed subject matter provides compositions and methods for the expression of CRISPR guide RNAs using the H1 promoter. In particular, compositions and methods are provided for the use of the H1 promoter to express CRISPR guide RNA (gRNA) with altered specificity of the 5′ nucleotide, as well as use of the H1 promoter sequence as a bidirectional promoter to express Cas9 nuclease and the gRNA simultaneously. Compositions and methods are also provided for the expression and regulation of gRNA expression in vivo through the use of RNA ribozymes and regulatable aptazymes.

公开(公告)号：US20190314521A1

公开(公告)日：2019-10-17

申请号：US16315458

申请日：2017-07-05

申请人： The Johns Hopkins University

发明人： Vinod Jaskula-Ranga ,  Donald Zack ,  Derek Welsbie

IPC分类号： A61K48/00 ,  A61K9/00 ,  C12N15/86

摘要： The presently disclosed subject matter provides compositions and methods comprising improvements of a CRISPR system (e.g. CRISPR associated (Cas) 9 (CRISPR-Cas9, non-Cas9 CRISPR systems). Such compositions may comprise modifications to the H1 promoter region, addition of 5′UTR modifications, different orthologous sequences of the H1 promoter, novel compact bidirectional promoter sequences with both pol II and pol III activity, addition of Kozak consensus sequences, termination sequences, addition of conditional pol II/pol III bidirectional promoter expression, addition of a donor template sequence for correcting mutations, or combinations thereof. Other aspects of the invention relate to modifications to Cas9 through post-transcriptional cell-cycle regulation fusions, engineered partial target sites such that the nuclease can bind without DNA cleavage, auto-regulation sites, and N-terminal modifications to modulate half-life.

公开(公告)号：US20180200388A1

公开(公告)日：2018-07-19

申请号：US15864715

申请日：2018-01-08

申请人： The Johns Hopkins University

发明人： Vinod Jaskula-Ranga ,  Donald Zack

IPC分类号： A61K48/00 ,  C12N15/113 ,  C12N15/63 ,  C12N15/85 ,  C12N15/90

CPC分类号： A61K48/0066 ,  C07K2319/10 ,  C07K2319/60 ,  C12N15/111 ,  C12N15/1138 ,  C12N15/63 ,  C12N15/85 ,  C12N15/86 ,  C12N15/907 ,  C12N2310/10 ,  C12N2310/20 ,  C12N2330/51 ,  C12N2750/14133 ,  C12N2750/14143 ,  C12N2830/205 ,  C12N2830/85

摘要： The presently disclosed subject matter provides compositions and methods for the expression of CRISPR guide RNAs using the H1 promoter. In particular, compositions and methods are provided for the use of the H1 promoter to express CRISPR guide RNA (gRNA) with altered specificity of the 5′ nucleotide, as well as use of the H1 promoter sequence as a bidirectional promoter to express Cas9 nuclease and the gRNA simultaneously. Compositions and methods are also provided for the expression and regulation of gRNA expression in vivo through the use of RNA ribozymes and regulatable aptazymes.

公开(公告)号：US20240336934A1

公开(公告)日：2024-10-10

申请号：US18380920

申请日：2023-10-17

申请人： The Johns Hopkins University

发明人： Vinod Jaskula-Ranga ,  Donald Zack ,  Fred Bunz ,  Derek Welsbie

IPC分类号： C12N15/86 ,  A61K38/46 ,  A61K48/00 ,  C12N9/22 ,  C12N15/113 ,  C12N15/90

CPC分类号： C12N15/86 ,  A61K38/465 ,  A61K48/005 ,  A61K48/0075 ,  C12N9/22 ,  C12N15/113 ,  C12N15/907 ,  C12N2310/121 ,  C12N2310/20 ,  C12N2310/3519 ,  C12N2320/35 ,  C12N2330/51 ,  C12N2750/14343 ,  C12N2830/205

摘要： Described herein are methods for treating a retinal degeneration in a subject, such as Leber's congenital amaurosis (LCA), retinitis pigmentosa (RP), and glaucoma. Also provided herein are methods of altering expression of one or more gene products in a cell, such as a retinal ganglion cell. Such methods may comprise utilizing a modified nuclease system, such as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system comprising a bidirectional HI promoter and gRNAs directed to retinal degeneration related genes, packaged in a single, compact adeno-associated virus (AAV) particle.

公开(公告)号：US20200080108A1

公开(公告)日：2020-03-12

申请号：US16315462

申请日：2017-07-05

申请人： The Johns Hopkins University

发明人： Vinod Jaskula-Ranga ,  Donald Zack ,  Fred Bunz ,  Derek Welsbie

IPC分类号： C12N15/86 ,  A61K38/46 ,  A61K48/00 ,  C12N15/113 ,  C12N15/90 ,  C12N9/22

摘要： Described herein are methods for treating a retinal degeneration in a subject, such as Leber's congenital amaurosis (LCA), retinitis pigmentosa (RP), and glaucoma. Also provided herein are methods of altering expression of one or more gene products in a cell, such as a retinal ganglion cell. Such methods may comprise utilizing a modified nuclease system, such as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system comprising a bidirectional HI promoter and gRNAs directed to retinal degeneration related genes, packaged in a single, compact adeno-associated virus (AAV) particle.

公开(公告)号：US10369234B2

公开(公告)日：2019-08-06

申请号：US15864715

申请日：2018-01-08

申请人： The Johns Hopkins University

发明人： Vinod Jaskula-Ranga ,  Donald Zack

IPC分类号： A61K48/00 ,  C12N15/63 ,  C12N15/85 ,  C12N15/90 ,  C12N15/113 ,  C12N15/11 ,  C12N15/86

摘要： The presently disclosed subject matter provides compositions and methods for the expression of CRISPR guide RNAs using the H1 promoter. In particular, compositions and methods are provided for the use of the H1 promoter to express CRISPR guide RNA (gRNA) with altered specificity of the 5′ nucleotide, as well as use of the H1 promoter sequence as a bidirectional promoter to express Cas9 nuclease and the gRNA simultaneously. Compositions and methods are also provided for the expression and regulation of gRNA expression in vivo through the use of RNA ribozymes and regulatable aptazymes.

公开(公告)号：US09907863B2

公开(公告)日：2018-03-06

申请号：US14951240

申请日：2015-11-24

申请人： THE JOHNS HOPKINS UNIVERSITY

发明人： Vinod Jaskula-Ranga ,  Donald Zack

IPC分类号： A61K48/00 ,  C12N15/63 ,  C12N15/85 ,  C12N15/90 ,  C12N15/113 ,  C12N15/86

CPC分类号： A61K48/0066 ,  C07K2319/10 ,  C07K2319/60 ,  C12N15/111 ,  C12N15/1138 ,  C12N15/63 ,  C12N15/85 ,  C12N15/907 ,  C12N2310/10 ,  C12N2310/20 ,  C12N2330/51 ,  C12N2750/14133 ,  C12N2750/14143 ,  C12N2830/85

摘要： The presently disclosed subject matter provides compositions and methods for the expression of CRISPR guide RNAs using the H1 promoter. In particular, compositions and methods are provided for the use of the H1 promoter to express CRISPR guide RNA (gRNA) with altered specificity of the 5′ nucleotide, as well as use of the H1 promoter sequence as a bidirectional promoter to express Cas9 nuclease and the gRNA simultaneously. Compositions and methods are also provided for the expression and regulation of gRNA expression in vivo through the use of RNA ribozymes and regulatable aptazymes.

公开(公告)号：US11896679B2

公开(公告)日：2024-02-13

申请号：US16851636

申请日：2020-04-17

申请人： The Johns Hopkins University

发明人： Vinod Jaskula-Ranga ,  Donald Zack

IPC分类号： A61K48/00 ,  C12N15/85 ,  C12N15/113 ,  C12N15/63 ,  C12N15/90 ,  C12N15/11 ,  C12N15/861 ,  C12N15/86 ,  C12Q1/68

CPC分类号： A61K48/0066 ,  C12N15/111 ,  C12N15/1138 ,  C12N15/63 ,  C12N15/85 ,  C12N15/86 ,  C12N15/8616 ,  C12N15/907 ,  C07K2319/10 ,  C07K2319/60 ,  C12N2310/10 ,  C12N2310/20 ,  C12N2330/51 ,  C12N2750/14133 ,  C12N2750/14143 ,  C12N2830/205 ,  C12N2830/85

摘要： The presently disclosed subject matter provides compositions and methods for the expression of CRISPR guide RNAs using the H1 promoter. In particular, compositions and methods are provided for the use of the H1 promoter to express CRISPR guide RNA (gRNA) with altered specificity of the 5′ nucleotide, as well as use of the H1 promoter sequence as a bidirectional promoter to express Cas9 nuclease and the gRNA simultaneously. Compositions and methods are also provided for the expression and regulation of gRNA expression in vivo through the use of RNA ribozymes and regulatable aptazymes.