公开(公告)号：US20220177913A1

公开(公告)日：2022-06-09

申请号：US17672744

申请日：2022-02-16

Applicant: President and Fellows of Harvard College

Inventor： George M. Church ,  Prashant G. Mali ,  Luhan Yang

IPC: C12N15/85 ,  C12N15/10 ,  C12N15/63 ,  C12N15/90 ,  C12N15/01 ,  C12N15/81 ,  C12N15/82 ,  C12N15/87 ,  C12N9/22

Abstract: A method of altering a eukaryotic cell is provided including transfecting the eukaryotic cell with a nucleic acid encoding RNA complementary to genomic DNA of the eukaryotic cell, transfecting the eukaryotic cell with a nucleic acid encoding an enzyme that interacts with the RNA and cleaves the genomic DNA in a site specific manner, wherein the cell expresses the RNA and the enzyme, the RNA binds to complementary genomic DNA and the enzyme cleaves the genomic DNA in a site specific manner.

公开(公告)号：US20210222193A1

公开(公告)日：2021-07-22

申请号：US17186139

申请日：2021-02-26

Applicant: President and Fellows of Harvard College

Inventor： George M. Church ,  Prashant G. Mali ,  Luhan Yang

IPC: C12N15/85 ,  C12N15/10 ,  C12N15/63 ,  C12N15/90 ,  C12N15/01 ,  C12N15/81 ,  C12N15/82 ,  C12N15/87 ,  C12N9/22

Abstract: A method of altering a eukaryotic cell is provided including transfecting the eukaryotic cell with a nucleic acid encoding RNA complementary to genomic DNA of the eukaryotic cell, transfecting the eukaryotic cell with a nucleic acid encoding an enzyme that interacts with the RNA and cleaves the genomic DNA in a site specific manner, wherein the cell expresses the RNA and the enzyme, the RNA binds to complementary genomic DNA and the enzyme cleaves the genomic DNA in a site specific manner.

公开(公告)号：US20200299732A1

公开(公告)日：2020-09-24

申请号：US16851360

申请日：2020-04-17

Applicant: President and Fellows of Harvard College

Inventor： George M. Church ,  Prashant G. Mali ,  Kevin M. Esvelt

IPC: C12N15/90 ,  C12N15/113 ,  C12N15/10 ,  C12N15/63 ,  C12N9/22 ,  C12N15/11

Abstract: Methods of modulating expression of a target nucleic acid in a cell are provided including introducing into the cell a first foreign nucleic acid encoding one or more RNAs complementary to DNA, wherein the DNA includes the target nucleic acid, introducing into the cell a second foreign nucleic acid encoding a nuclease-null Cas9 protein that binds to the DNA and is guided by the one or more RNAs, introducing into the cell a third foreign nucleic acid encoding a transcriptional regulator protein or domain, wherein the one or more RNAs, the nuclease-null Cas9 protein, and the transcriptional regulator protein or domain are expressed, wherein the one or more RNAs, the nuclease-null Cas9 protein and the transcriptional regulator protein or domain co-localize to the DNA and wherein the transcriptional regulator protein or domain regulates expression of the target nucleic acid.

公开(公告)号：US10435708B2

公开(公告)日：2019-10-08

申请号：US15042515

申请日：2016-02-12

Applicant: President and Fellows of Harvard College

Inventor： Prashant G. Mali ,  George M. Church ,  Luhan Yang

IPC: C12Q1/68 ,  C12N15/00 ,  C12N15/85 ,  C12N15/10 ,  C12N15/90 ,  C12N15/63 ,  C12N15/01 ,  C12N15/81 ,  C12N15/82 ,  C12N15/87 ,  C12N9/22

Abstract: A method of altering a eukaryotic cell is provided including transfecting the eukaryotic cell with a nucleic acid encoding RNA complementary to genomic DNA of the eukaryotic cell, transfecting the eukaryotic cell with a nucleic acid encoding an enzyme that interacts with the RNA and cleaves the genomic DNA in a site specific manner, wherein the cell expresses the RNA and the enzyme, the RNA binds to complementary genomic DNA and the enzyme cleaves the genomic DNA in a site specific manner.

公开(公告)号：US20190264202A1

公开(公告)日：2019-08-29

申请号：US16346569

申请日：2017-11-01

Applicant: President and Fellows of Harvard College

Inventor： George M. Church ,  Reza Kalhor ,  Prashant G. Mali

IPC: C12N15/11 ,  C12Q1/6853 ,  C12N15/10

Abstract: A functional engineered guide RNA sequence is provided including a spacer sequence and a scaffold sequence, wherein the scaffold sequence includes a primer binding site for reverse transcription.

公开(公告)号：US20190249193A1

公开(公告)日：2019-08-15

申请号：US16397213

申请日：2019-04-29

Applicant: President and Fellows of Harvard College

Inventor： George M. Church ,  Prashant G. Mali ,  Luhan Yang

IPC: C12N15/85 ,  C12N15/10 ,  C12N15/82 ,  C12N15/81 ,  C12N9/22 ,  C12N15/90 ,  C12N15/63 ,  C12N15/87 ,  C12N15/01

CPC classification number: C12N15/85 ,  C12N9/22 ,  C12N15/01 ,  C12N15/10 ,  C12N15/102 ,  C12N15/1024 ,  C12N15/63 ,  C12N15/81 ,  C12N15/8201 ,  C12N15/87 ,  C12N15/90 ,  C12N15/907 ,  C12N2310/20 ,  C12N2800/80 ,  C12N2810/55 ,  C12Y301/00

Abstract: A method of altering a eukaryotic cell is provided including transfecting the eukaryotic cell with a nucleic acid encoding RNA complementary to genomic DNA of the eukaryotic cell, transfecting the eukaryotic cell with a nucleic acid encoding an enzyme that interacts with the RNA and cleaves the genomic DNA in a site specific manner, wherein the cell expresses the RNA and the enzyme, the RNA binds to complementary genomic DNA and the enzyme cleaves the genomic DNA in a site specific manner.

公开(公告)号：US10329587B2

公开(公告)日：2019-06-25

申请号：US14903728

申请日：2014-07-08

Applicant: President and Fellows of Harvard College

Inventor： George M. Church ,  Kevin M. Esvelt ,  Prashant G. Mali

IPC: C12N15/113 ,  C12N9/22 ,  C12N15/90 ,  C12N15/85 ,  C07K2/00 ,  C12N15/11

Abstract: Methods of modulating expression of a target nucleic acid in a cell are provided including use of multiple orthogonal Cas9 proteins to simultaneously and independently regulate corresponding genes or simultaneously and independently edit corresponding genes.

公开(公告)号：US09267135B2

公开(公告)日：2016-02-23

申请号：US14319289

申请日：2014-06-30

Applicant: President and Fellows of Harvard College

Inventor： George M. Church ,  Prashant G. Mali ,  Kevin M. Esvelt

IPC: C12N15/63 ,  C12N15/113 ,  C12N15/66 ,  C12N15/85 ,  C12N15/01 ,  C12N15/87 ,  C12N15/10

CPC classification number: C12N15/907 ,  C12N9/22 ,  C12N15/102 ,  C12N15/11 ,  C12N15/113 ,  C12N15/635 ,  C12N2310/20 ,  C12N2310/3513 ,  C12Y301/00

Abstract: Methods of modulating expression of a target nucleic acid in a cell are provided including introducing into the cell a first foreign nucleic acid encoding one or more RNAs complementary to DNA, wherein the DNA includes the target nucleic acid, introducing into the cell a second foreign nucleic acid encoding a nuclease-null Cas9 protein that binds to the DNA and is guided by the one or more RNAs, introducing into the cell a third foreign nucleic acid encoding a transcriptional regulator protein or domain, wherein the one or more RNAs, the nuclease-null Cas9 protein, and the transcriptional regulator protein or domain are expressed, wherein the one or more RNAs, the nuclease-null Cas9 protein and the transcriptional regulator protein or domain co-localize to the DNA and wherein the transcriptional regulator protein or domain regulates expression of the target nucleic acid.

Abstract translation: 提供了调节细胞中靶核酸表达的方法，包括向细胞中引入编码与DNA互补的一个或多个RNA的第一外源核酸，其中DNA包括靶核酸，将第二外源核酸 编码与所述DNA结合并由所述一个或多个RNA引导的核酸酶 - 无效Cas9蛋白的酸，其将所述一个或多个RNA引入所述细胞中，所述第三外源核酸编码转录调节蛋白或结构域，其中所述核酸酶 - 表达无效Cas9蛋白和转录调节蛋白或结构域，其中一个或多个RNA，核酸酶无效Cas9蛋白和转录调节蛋白或结构域共定位于DNA，并且其中转录调节蛋白或结构域调节表达 的靶核酸。

公开(公告)号：US09023649B2

公开(公告)日：2015-05-05

申请号：US14319100

申请日：2014-06-30

Applicant: President and Fellows of Harvard College

Inventor： Prashant G. Mali ,  George M. Church ,  Luhan Yang

IPC: C12N15/85 ,  C12N9/14 ,  C12N9/22 ,  C07H21/02 ,  C07H21/04 ,  C12N15/87 ,  C12N15/01 ,  C12N15/81 ,  C12N15/82

CPC classification number: C12N15/85 ,  C12N9/22 ,  C12N15/01 ,  C12N15/10 ,  C12N15/102 ,  C12N15/1024 ,  C12N15/63 ,  C12N15/81 ,  C12N15/8201 ,  C12N15/87 ,  C12N15/90 ,  C12N15/907 ,  C12N2800/80 ,  C12N2810/55 ,  C12Y301/00

Abstract: A method of altering a eukaryotic cell is provided including transfecting the eukaryotic cell with a nucleic acid encoding RNA complementary to genomic DNA of the eukaryotic cell, transfecting the eukaryotic cell with a nucleic acid encoding an enzyme that interacts with the RNA and cleaves the genomic DNA in a site specific manner, wherein the cell expresses the RNA and the enzyme, the RNA binds to complementary genomic DNA and the enzyme cleaves the genomic DNA in a site specific manner.

Abstract translation: 提供了改变真核细胞的方法，包括用编码与真核细胞的基因组DNA互补的RNA的核酸转染真核细胞，用编码与RNA相互作用并切割基因组DNA的酶的核酸转染真核细胞 以位点特异性方式，其中细胞表达RNA和酶，RNA结合互补基因组DNA，酶以位点特异性方式切割基因组DNA。