JPWO2018232356A5 - - Google Patents
Download PDFInfo
- Publication number
- JPWO2018232356A5 JPWO2018232356A5 JP2019569305A JP2019569305A JPWO2018232356A5 JP WO2018232356 A5 JPWO2018232356 A5 JP WO2018232356A5 JP 2019569305 A JP2019569305 A JP 2019569305A JP 2019569305 A JP2019569305 A JP 2019569305A JP WO2018232356 A5 JPWO2018232356 A5 JP WO2018232356A5
- Authority
- JP
- Japan
- Prior art keywords
- dna template
- cell
- rnp
- cells
- primary
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Description
いくつかの態様において、RNP-DNA鋳型複合体は約1×105個～約2×106個の細胞に導入される。いくつかの態様において、細胞は初代造血細胞または初代造血幹細胞である。いくつかの態様において、初代造血細胞は免疫細胞である。いくつかの態様において、免疫細胞はT細胞である。いくつかの態様において、T細胞は制御性T細胞、エフェクターT細胞またはナイーブT細胞である。いくつかの態様において、T細胞はCD8+T細胞である。いくつかの態様において、T細胞はCD4+CD8+T細胞である。
[本発明1001]
（a）Cas9リボ核タンパク質複合体（RNP）-DNA鋳型複合体を提供する工程であって、該RNP-DNA鋳型複合体が、
（i）RNPがCas9ヌクレアーゼドメインおよびガイドRNAを含み、該ガイドRNAが細胞のゲノムの標的領域に特異的にハイブリダイズし、該Cas9ヌクレアーゼドメインが該標的領域を切断して該細胞の該ゲノム中に挿入部位を生じさせる、該RNPと；
（ii）二本鎖または一本鎖のDNA鋳型であって、該DNA鋳型のサイズが約200ヌクレオチドよりも大きく、該DNA鋳型の5'および3'末端が、該挿入部位に隣接するゲノム配列と相同であるヌクレオチド配列を含む、DNA鋳型と
を含み、該複合体におけるRNP：DNA鋳型のモル比が約3:1～約100:1である、工程；ならびに
（b）該RNP-DNA鋳型複合体を細胞に導入する工程
を含む、細胞のゲノムを編集する方法。
[本発明1002]
RNP-DNA鋳型複合体が、RNPをDNA鋳型とともに約20～25℃の温度で約1分未満～約30分の間インキュベートすることによって形成される、本発明1001の方法。
[本発明1003]
DNA鋳型が直鎖状DNA鋳型である、本発明1001または1002の方法。
[本発明1004]
DNA鋳型が一本鎖DNA鋳型である、本発明1001～1003のいずれかの方法。
[本発明1005]
DNA鋳型が純粋な一本鎖DNA鋳型である、本発明1001～1003のいずれかの方法。
[本発明1006]
RNP-DNA鋳型複合体と細胞とが、該RNP-DNA鋳型複合体を該細胞に導入する前に混合される、本発明1001～1005のいずれかの方法。
[本発明1007]
RNPがCas9ヌクレアーゼを含む、本発明1001～1006のいずれかの方法。
[本発明1008]
RNPがCas9ニッカーゼを含む、本発明1001～1006のいずれかの方法。
[本発明1009]
RNP-DNA鋳型複合体が少なくとも2つの構造的に異なるRNP複合体を含む、本発明1001～1008のいずれかの方法。
[本発明1010]
少なくとも2つの構造的に異なるRNP複合体が構造的に異なるガイドRNAを含む、本発明1009の方法。
[本発明1011]
構造的に異なるRNP複合体それぞれがCas9ニッカーゼを含み、構造的に異なるガイドRNAが標的領域の反対の鎖にハイブリダイズする、本発明1010の方法。
[本発明1012]
少なくとも2つの構造的に異なるRNP複合体が構造的に異なるCas9ヌクレアーゼドメインを含む、本発明1009の方法。
[本発明1013]
前記導入する工程がエレクトロポレーションを含む、本発明1001～1012のいずれかの方法。
[本発明1014]
RNP：DNA鋳型のモル比が約5：1～約15：1である、本発明1001～1013のいずれかの方法。
[本発明1015]
RNP：DNA鋳型のモル比が約5：1～約10：1である、本発明1001～1014のいずれかの方法。
[本発明1016]
RNP：DNA鋳型のモル比が約8：1～約12：1である、本発明1001～1015のいずれかの方法。
[本発明1017]
DNA鋳型のサイズが約1kbよりも大きい、本発明1001～1016のいずれかの方法。
[本発明1018]
DNA鋳型が約2.5pM～約25pMの濃度である、本発明1001～1017のいずれかの方法。
[本発明1019]
DNA鋳型の量が約1μg～約10μgである、本発明1001～1018のいずれかの方法。
[本発明1020]
細胞が初代造血細胞または初代造血幹細胞である、本発明1001～1019のいずれかの方法。
[本発明1021]
RNP-DNA鋳型複合体が約1×10
5
個～約2×10
6
個の細胞に導入される、本発明1001～1020のいずれかの方法。
[本発明1022]
細胞が初代造血細胞である、本発明1001～1021のいずれかの方法。
[本発明1023]
初代造血細胞が免疫細胞である、本発明1022の方法。
[本発明1024]
免疫細胞がT細胞である、本発明1023の方法。
[本発明1025]
T細胞が制御性T細胞、エフェクターT細胞またはナイーブT細胞である、本発明1024の方法。
[本発明1026]
制御性T細胞、エフェクターT細胞またはナイーブT細胞がCD4
+
T細胞である、本発明1025の方法。
[本発明1027]
T細胞がCD8
+
T細胞である、本発明1024の方法。
[本発明1028]
T細胞がCD4
+
CD8
+
T細胞である、本発明1024の方法。
In some embodiments, the RNP-DNA template complex is introduced into about 1 × 10 5 to about 2 × 10 6 cells. In some embodiments, the cell is a primary hematopoietic cell or a primary hematopoietic stem cell. In some embodiments, the primary hematopoietic cells are immune cells. In some embodiments, the immune cell is a T cell. In some embodiments, the T cell is a regulatory T cell, an effector T cell or a naive T cell. In some embodiments, the T cell is a CD8 + T cell. In some embodiments, the T cell is a CD4 + CD8 + T cell.
[Invention 1001]
(A) A step of providing a Cas9 ribonuclear protein complex (RNP) -DNA template complex, wherein the RNP-DNA template complex is used.
(I) The RNP contains a Cas9 nuclease domain and a guide RNA, the guide RNA specifically hybridizes to the target region of the cell's genome, and the Cas9 nuclease domain cleaves the target region into the genome of the cell. With the RNA, which gives rise to an insertion site in
(Ii) A double- or single-stranded DNA template in which the size of the DNA template is greater than about 200 nucleotides, and the 5'and 3'ends of the DNA template are genomic sequences flanking the insertion site. With a DNA template containing a nucleotide sequence homologous to
The molar ratio of RNP: DNA template in the complex is from about 3: 1 to about 100: 1, steps;
(B) Step of introducing the RNP-DNA template complex into cells
How to edit the genome of a cell, including.
[Invention 1002]
The method of the present invention 1001 wherein the RNP-DNA template complex is formed by incubating RNP with the DNA template at a temperature of about 20-25 ° C for less than about 1 minute to about 30 minutes.
[Invention 1003]
The method of the present invention 1001 or 1002, wherein the DNA template is a linear DNA template.
[Invention 1004]
The method according to any one of 1001 to 1003 of the present invention, wherein the DNA template is a single-stranded DNA template.
[Invention 1005]
The method of any of 1001-1003 of the present invention, wherein the DNA template is a pure single-stranded DNA template.
[Invention 1006]
The method of any of 1001 to 1005 of the present invention, wherein the RNP-DNA template complex and the cell are mixed prior to introducing the RNP-DNA template complex into the cell.
[Invention 1007]
The method of any of 1001-1006 of the present invention, wherein the RNP comprises Cas9 nuclease.
[Invention 1008]
The method of any of 1001-1006 of the present invention, wherein the RNP comprises Cas9 nickase.
[Invention 1009]
The method of any of 1001-1008 of the present invention, wherein the RNP-DNA template complex comprises at least two structurally distinct RNP complexes.
[Invention 1010]
The method of the present invention 1009, wherein at least two structurally distinct RNP complexes contain structurally distinct guide RNAs.
[Invention 1011]
The method of the invention 1010, wherein each of the structurally distinct RNP complexes contains Cas9 nickase and the structurally distinct guide RNA hybridizes to the opposite strand of the target region.
[Invention 1012]
The method of the present invention 1009, wherein at least two structurally distinct RNP complexes contain structurally distinct Cas9 nuclease domains.
[Invention 1013]
The method of any of 1001 to 1012 of the present invention, wherein the process of introduction comprises electroporation.
[Invention 1014]
The method according to any one of 1001 to 1013 of the present invention, wherein the molar ratio of the RNP: DNA template is about 5: 1 to about 15: 1.
[Invention 1015]
The method according to any one of 1001 to 1014 of the present invention, wherein the molar ratio of the RNP: DNA template is about 5: 1 to about 10: 1.
[Invention 1016]
The method according to any one of 1001 to 1015 of the present invention, wherein the molar ratio of the RNP: DNA template is about 8: 1 to about 12: 1.
[Invention 1017]
The method of any of 1001-1016 of the present invention, wherein the size of the DNA template is greater than about 1 kb.
[Invention 1018]
The method of any of 1001-1017 of the present invention, wherein the DNA template has a concentration of about 2.5 pM to about 25 pM.
[Invention 1019]
The method of any of 1001-1018 of the present invention, wherein the amount of DNA template is from about 1 μg to about 10 μg.
[Invention 1020]
The method of any of 1001-1019 of the present invention, wherein the cells are primary hematopoietic cells or primary hematopoietic stem cells.
[Invention 1021]
The method of any of 1001 to 1020 of the present invention, wherein the RNP-DNA template complex is introduced into about 1 × 10 5 to about 2 × 10 6 cells.
[Invention 1022]
The method of any of 1001 to 1021 of the present invention, wherein the cell is a primary hematopoietic cell.
[Invention 1023]
The method of 1022 of the present invention, wherein the primary hematopoietic cells are immune cells.
[Invention 1024]
The method of the present invention 1023, wherein the immune cells are T cells.
[Invention 1025]
The method of the invention 1024, wherein the T cells are regulatory T cells, effector T cells or naive T cells.
[Invention 1026]
The method of the invention 1025, wherein the regulatory T cells, effector T cells or naive T cells are CD4 + T cells.
[Invention 1027]
The method of the present invention 1024, wherein the T cells are CD8 + T cells.
[Invention 1028]
The method of the present invention 1024, wherein the T cells are CD4 + CD8 + T cells.
Claims (35)
（i）RNPがCas9ヌクレアーゼドメインおよびガイドRNAを含み、該ガイドRNAが細胞のゲノムの標的領域に特異的にハイブリダイズし、該Cas9ヌクレアーゼドメインが該標的領域を切断して該細胞の該ゲノム中に挿入部位を生じさせる、該RNPと；
（ii）二本鎖または一本鎖のDNA鋳型であって、該DNA鋳型のサイズが約200ヌクレオチドよりも大きく、該DNA鋳型の5'および3'末端が、該挿入部位に隣接するゲノム配列と相同であるヌクレオチド配列を含む、DNA鋳型と
を含み、該複合体におけるRNP：DNA鋳型のモル比が約3:1～約100:1である、工程；ならびに
（b）該RNP-DNA鋳型複合体を細胞に導入する工程であって、該RNP-DNA鋳型複合体と該細胞とが、該RNP-DNA鋳型複合体を該細胞に導入する前に混合される、工程
を含む、細胞のゲノムを編集するインビトロまたはエクスビボ方法。 (A) A step of providing a Cas9 ribonuclear protein complex (RNP) -DNA template complex, wherein the RNP-DNA template complex is used.
(I) The RNP contains a Cas9 nuclease domain and a guide RNA, the guide RNA specifically hybridizes to the target region of the cell's genome, and the Cas9 nuclease domain cleaves the target region into the genome of the cell. With the RNA, which gives rise to an insertion site in
(Ii) A double- or single-stranded DNA template in which the size of the DNA template is greater than about 200 nucleotides, and the 5'and 3'ends of the DNA template are genomic sequences flanking the insertion site. The RNP: DNA template has a molar ratio of about 3: 1 to about 100: 1 in the complex, comprising a DNA template comprising a nucleotide sequence homologous to, and (b) the RNP-DNA template. A step of introducing the complex into a cell, wherein the RNP-DNA template complex and the cell are mixed before the RNP-DNA template complex is introduced into the cell.
In vitro or exvivo methods for editing the genome of a cell, including.
（b）DNA鋳型が一本鎖DNA鋳型、任意で純粋な一本鎖DNA鋳型である、または
（c）（i）DNA鋳型が細胞のゲノムに非ウイルス的に挿入されるか、もしくは（ii）DNA鋳型がウイルスベクターの非存在下で細胞のゲノムに挿入される、
請求項1または2記載の方法。 (A) The DNA template is a linear DNA template,
( B) The DNA template is a single-stranded DNA template, optionally a pure single-stranded DNA template, or
(C) (i) The DNA template is non-virally inserted into the cell's genome, or (ii) the DNA template is inserted into the cell's genome in the absence of a viral vector.
The method of claim 1 or 2.
（b）RNPがCas9ニッカーゼを含む、
請求項1～3のいずれか一項記載の方法。 (A) RNP contains Cas9 nuclease, or
(B) RNP contains Cas9 nickase,
The method according to any one of claims 1 to 3 .
（b）少なくとも2つの構造的に異なるRNP複合体が構造的に異なるCas9ヌクレアーゼドメインを含む、
請求項5記載の方法。 (A) At least two structurally different RNP complexes contain structurally different guide RNAs, optionally each structurally different RNP complex contains Cas9 nickase, and structurally different guide RNAs are targeted regions. Hybridizes to the opposite chain of, or
(B) At least two structurally distinct RNP complexes contain structurally distinct Cas9 nuclease domains.
The method according to claim 5 .
（b）RNP：DNA鋳型のモル比が約5：1～約10：1である、または
（c）RNP：DNA鋳型のモル比が約8：1～約12：1である、
請求項1～7のいずれか一項記載の方法。 (A) The molar ratio of RNP: DNA template is about 5: 1 to about 15: 1.
(B) The molar ratio of RNP: DNA template is about 5: 1 to about 10: 1, or
(C) The molar ratio of RNP: DNA template is about 8: 1 to about 12: 1,
The method according to any one of claims 1 to 7 .
（b）DNA鋳型が約2.5pM～約25pMの濃度である、および/または
（c）DNA鋳型の量が約1μg～約10μgである、
請求項1～8のいずれか一項記載の方法。 (A) The size of the DNA template is larger than about 1 kb,
(B) The DNA template has a concentration of about 2.5 pM to about 25 pM, and / or
(C) The amount of DNA template is about 1 μg to about 10 μg,
The method according to any one of claims 1 to 8 .
（b）T細胞がCD8 + T細胞である、または
（c）T細胞がCD4 + CD8 + T細胞である、
請求項13記載の方法。 (A) T cells are regulatory T cells, effector T cells , or naive T cells, and optionally regulatory T cells, effector T cells, or naive T cells are CD4 + T cells. ,
(B) T cells are CD8 + T cells, or
(C) T cells are CD4 + CD8 + T cells,
13. The method of claim 13 .
リボ核タンパク質複合体（RNP）-DNA鋳型複合体を提供する工程であって、該RNPがヌクレアーゼドメインおよびガイドRNAを含み、該DNA鋳型のサイズが約200ヌクレオチド以上であり、該DNA鋳型の5'および3'末端が、初代免疫細胞のゲノム中の挿入部位に隣接するゲノム配列と相同であるヌクレオチド配列を含む、工程；A step of providing a ribonuclear protein complex (RNP) -DNA template complex, wherein the RNP contains a nuclease domain and a guide RNA, the size of the DNA template is about 200 nucleotides or more, and 5 of the DNA template. A step comprising a nucleotide sequence in which the'and 3'ends are homologous to the genomic sequence flanking the insertion site in the genome of the primary immune cell;
該RNP-DNA鋳型複合体を初代免疫細胞に非ウイルス的に導入する工程であって、該ガイドRNAが初代免疫細胞のゲノムの標的領域に特異的にハイブリダイズし、該ヌクレアーゼドメインが該標的領域を切断して該初代免疫細胞のゲノム中に挿入部位を生じさせる、工程；ならびにIn the step of non-virally introducing the RNP-DNA template complex into a primary immune cell, the guide RNA specifically hybridizes to the target region of the genome of the primary immune cell, and the nuclease domain is the target region. To generate an insertion site in the genome of the primary immune cell, step;
該初代免疫細胞のゲノム中の挿入部位にDNA鋳型を挿入することにより、該初代免疫細胞を編集する工程。A step of editing a primary immune cell by inserting a DNA template into the insertion site in the genome of the primary immune cell.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2023076182A JP2023100828A (en) | 2017-06-15 | 2023-05-02 | Targeted non-viral dna insertions |
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201762520117P | 2017-06-15 | 2017-06-15 | |
| US62/520,117 | 2017-06-15 | ||
| US201762552180P | 2017-08-30 | 2017-08-30 | |
| US62/552,180 | 2017-08-30 | ||
| PCT/US2018/037919 WO2018232356A1 (en) | 2017-06-15 | 2018-06-15 | Targeted non-viral dna insertions |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2023076182A Division JP2023100828A (en) | 2017-06-15 | 2023-05-02 | Targeted non-viral dna insertions |
Publications (4)
| Publication Number | Publication Date |
|---|---|
| JP2020524998A JP2020524998A (en) | 2020-08-27 |
| JP2020524998A5 JP2020524998A5 (en) | 2021-07-29 |
| JPWO2018232356A5 true JPWO2018232356A5 (en) | 2022-02-07 |
| JP7275054B2 JP7275054B2 (en) | 2023-05-17 |
Family
ID=64659467
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2019569305A Active JP7275054B2 (en) | 2017-06-15 | 2018-06-15 | Targeted non-viral DNA insertion |
| JP2023076182A Pending JP2023100828A (en) | 2017-06-15 | 2023-05-02 | Targeted non-viral dna insertions |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2023076182A Pending JP2023100828A (en) | 2017-06-15 | 2023-05-02 | Targeted non-viral dna insertions |
Country Status (12)
| Country | Link |
|---|---|
| US (4) | US20200362355A1 (en) |
| EP (1) | EP3638317A4 (en) |
| JP (2) | JP7275054B2 (en) |
| KR (2) | KR20200018645A (en) |
| CN (1) | CN111344020A (en) |
| AU (1) | AU2018283405A1 (en) |
| BR (1) | BR112019026625A2 (en) |
| CA (1) | CA3067382A1 (en) |
| IL (1) | IL271389A (en) |
| MX (1) | MX2019015188A (en) |
| SG (1) | SG11201912179SA (en) |
| WO (1) | WO2018232356A1 (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019084552A1 (en) | 2017-10-27 | 2019-05-02 | The Regents Of The University Of California | Targeted replacement of endogenous t cell receptors |
| GB2589246A (en) | 2018-05-16 | 2021-05-26 | Synthego Corp | Methods and systems for guide RNA design and use |
| EP3833680A4 (en) * | 2018-05-30 | 2022-05-25 | The Regents of the University of California | Gene editing of monogenic disorders in human hematopoietic stem cells -- correction of x-linked hyper-igm syndrome (xhim) |
| KR20230056026A (en) * | 2020-08-27 | 2023-04-26 | 티뮤니티 테라퓨틱스 인크. | Vector-Free Process for Manufacturing Engineered Immune Cells |
| CN114107292B (en) * | 2020-08-27 | 2024-03-12 | 阿思科力(苏州)生物科技有限公司 | Gene editing system and method for site-directed insertion of exogenous gene |
| CN113046381A (en) * | 2021-04-12 | 2021-06-29 | 南华大学 | Method for separating specific protein-DNA complex in organism, fusion protein and preparation method thereof |
| WO2023220207A2 (en) * | 2022-05-10 | 2023-11-16 | Editas Medicine, Inc. | Genome editing of cells |
| WO2024059618A2 (en) | 2022-09-13 | 2024-03-21 | Arsenal Biosciences, Inc. | Immune cells having co-expressed tgfbr shrnas |
| WO2024059824A2 (en) | 2022-09-16 | 2024-03-21 | Arsenal Biosciences, Inc. | Immune cells with combination gene perturbations |
Family Cites Families (95)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6534261B1 (en) | 1999-01-12 | 2003-03-18 | Sangamo Biosciences, Inc. | Regulation of endogenous gene expression in cells using zinc finger proteins |
| US20020064802A1 (en) | 2000-04-28 | 2002-05-30 | Eva Raschke | Methods for binding an exogenous molecule to cellular chromatin |
| GB0124391D0 (en) | 2001-10-11 | 2001-11-28 | Gene Expression Technologies L | Control of gene expression |
| US20050136040A1 (en) | 2001-10-11 | 2005-06-23 | Imperial College Innovations Limited | Control of gene expression using a complex of an oligonucleotide and a regulatory peptide |
| US20030232410A1 (en) | 2002-03-21 | 2003-12-18 | Monika Liljedahl | Methods and compositions for using zinc finger endonucleases to enhance homologous recombination |
| ITMI20030821A1 (en) | 2003-04-18 | 2004-10-19 | Internat Ct For Genetic En Gineering And | CHEMICAL POLYPEPTIDES AND THEIR USE. |
| US20060182736A1 (en) | 2003-06-10 | 2006-08-17 | Kim Jin-Soo | Transducible dna-binding proteins |
| US8133733B2 (en) | 2003-10-24 | 2012-03-13 | Gencia Corporation | Nonviral vectors for delivering polynucleotides to target tissues |
| WO2005123962A2 (en) | 2004-06-14 | 2005-12-29 | The University Of Texas At Austin | Gene targeting in eukaryotic cells by group ii intron ribonucleoprotein particles |
| US10022457B2 (en) | 2005-08-05 | 2018-07-17 | Gholam A. Peyman | Methods to regulate polarization and enhance function of cells |
| DK2336362T3 (en) | 2005-08-26 | 2019-01-21 | Dupont Nutrition Biosci Aps | USE OF CRISPR-ASSOCIATED GENES (CAS) |
| PL2049663T3 (en) | 2006-08-11 | 2015-08-31 | Dow Agrosciences Llc | Zinc finger nuclease-mediated homologous recombination |
| US20110239315A1 (en) | 2009-01-12 | 2011-09-29 | Ulla Bonas | Modular dna-binding domains and methods of use |
| JP2012525146A (en) | 2009-04-28 | 2012-10-22 | プレジデント アンド フェロウズ オブ ハーバード カレッジ | Overcharged protein for cell penetration |
| WO2011059836A2 (en) | 2009-10-29 | 2011-05-19 | Trustees Of Dartmouth College | T cell receptor-deficient t cell compositions |
| US8956828B2 (en) | 2009-11-10 | 2015-02-17 | Sangamo Biosciences, Inc. | Targeted disruption of T cell receptor genes using engineered zinc finger protein nucleases |
| SG181601A1 (en) | 2009-12-10 | 2012-07-30 | Univ Minnesota | Tal effector-mediated dna modification |
| CA2796600C (en) | 2010-04-26 | 2019-08-13 | Sangamo Biosciences, Inc. | Genome editing of a rosa locus using zinc-finger nucleases |
| US8876458B2 (en) | 2011-01-25 | 2014-11-04 | United Technologies Corporation | Blade outer air seal assembly and support |
| WO2013074916A1 (en) | 2011-11-18 | 2013-05-23 | Board Of Regents, The University Of Texas System | Car+ t cells genetically modified to eliminate expression of t- cell receptor and/or hla |
| GB201122458D0 (en) | 2011-12-30 | 2012-02-08 | Univ Wageningen | Modified cascade ribonucleoproteins and uses thereof |
| US20130236504A1 (en) | 2012-03-06 | 2013-09-12 | Medical University Of South Carolina | Delivery System for Enhancing Drug Efficacy |
| LT3401400T (en) | 2012-05-25 | 2019-06-10 | The Regents Of The University Of California | Methods and compositions for rna-directed target dna modification and for rna-directed modulation of transcription |
| DE202013012597U1 (en) | 2012-10-23 | 2017-11-21 | Toolgen, Inc. | A composition for cleaving a target DNA comprising a guide RNA specific for the target DNA and a Cas protein-encoding nucleic acid or Cas protein, and their use |
| EP3138911B1 (en) | 2012-12-06 | 2018-12-05 | Sigma Aldrich Co. LLC | Crispr-based genome modification and regulation |
| US8697359B1 (en) | 2012-12-12 | 2014-04-15 | The Broad Institute, Inc. | CRISPR-Cas systems and methods for altering expression of gene products |
| EP4234696A3 (en) | 2012-12-12 | 2023-09-06 | The Broad Institute Inc. | Crispr-cas component systems, methods and compositions for sequence manipulation |
| EP2931899A1 (en) | 2012-12-12 | 2015-10-21 | The Broad Institute, Inc. | Functional genomics using crispr-cas systems, compositions, methods, knock out libraries and applications thereof |
| CA3081054A1 (en) | 2012-12-17 | 2014-06-26 | President And Fellows Of Harvard College | Rna-guided human genome engineering |
| KR20140101203A (en) | 2013-02-08 | 2014-08-19 | 이정민 | Auto buffing machine |
| CA2906553C (en) | 2013-03-15 | 2022-08-02 | The General Hospital Corporation | Rna-guided targeting of genetic and epigenomic regulatory proteins to specific genomic loci |
| US9937207B2 (en) | 2013-03-21 | 2018-04-10 | Sangamo Therapeutics, Inc. | Targeted disruption of T cell receptor genes using talens |
| US20160053274A1 (en) | 2013-04-02 | 2016-02-25 | Bayer Cropscience Nv | Targeted genome engineering in eukaryotes |
| ES2883131T3 (en) | 2013-05-29 | 2021-12-07 | Cellectis | Methods for modifying T cells for immunotherapy using the RNA-guided CAS nuclease system |
| US9890393B2 (en) | 2013-05-29 | 2018-02-13 | Cellectis | Methods for engineering T cells for immunotherapy by using RNA-guided CAS nuclease system |
| US9526784B2 (en) | 2013-09-06 | 2016-12-27 | President And Fellows Of Harvard College | Delivery system for functional nucleases |
| US10822606B2 (en) | 2013-09-27 | 2020-11-03 | The Regents Of The University Of California | Optimized small guide RNAs and methods of use |
| CN105899665B (en) | 2013-10-17 | 2019-10-22 | 桑格摩生物科学股份有限公司 | The delivering method and composition being transformed for nuclease-mediated genome project |
| WO2015073867A1 (en) | 2013-11-15 | 2015-05-21 | The United States Of America, As Represented By The Secretary, Department Of Health & Human Services | Engineering neural stem cells using homologous recombination |
| US10787684B2 (en) * | 2013-11-19 | 2020-09-29 | President And Fellows Of Harvard College | Large gene excision and insertion |
| WO2015089354A1 (en) * | 2013-12-12 | 2015-06-18 | The Broad Institute Inc. | Compositions and methods of use of crispr-cas systems in nucleotide repeat disorders |
| WO2015089486A2 (en) | 2013-12-12 | 2015-06-18 | The Broad Institute Inc. | Systems, methods and compositions for sequence manipulation with optimized functional crispr-cas systems |
| KR20160089527A (en) | 2013-12-12 | 2016-07-27 | 더 브로드 인스티튜트, 인코퍼레이티드 | Delivery, use and therapeutic applications of the crispr-cas systems and compositions for genome editing |
| MX2016007327A (en) | 2013-12-12 | 2017-03-06 | Broad Inst Inc | Delivery, use and therapeutic applications of the crispr-cas systems and compositions for targeting disorders and diseases using particle delivery components. |
| WO2015086795A1 (en) | 2013-12-13 | 2015-06-18 | Cellectis | Cas9 nuclease platform for microalgae genome engineering |
| KR20220100078A (en) | 2014-01-31 | 2022-07-14 | 팩터 바이오사이언스 인크. | Methods and products for nucleic acid production and delivery |
| WO2015115903A1 (en) | 2014-02-03 | 2015-08-06 | Academisch Ziekenhuis Leiden H.O.D.N. Lumc | Site-specific dna break-induced genome editing using engineered nucleases |
| KR20230152175A (en) | 2014-04-18 | 2023-11-02 | 에디타스 메디신, 인코포레이티드 | Crispr-cas-related methods, compositions and components for cancer immunotherapy |
| KR101826904B1 (en) | 2014-08-06 | 2018-02-08 | 기초과학연구원 | Immune-compatible cells created by nuclease-mediated editing of genes encoding Human Leukocyte Antigens |
| WO2016036754A1 (en) | 2014-09-02 | 2016-03-10 | The Regents Of The University Of California | Methods and compositions for rna-directed target dna modification |
| WO2016049251A1 (en) | 2014-09-24 | 2016-03-31 | The Broad Institute Inc. | Delivery, use and therapeutic applications of the crispr-cas systems and compositions for modeling mutations in leukocytes |
| EP3204513A2 (en) | 2014-10-09 | 2017-08-16 | Life Technologies Corporation | Crispr oligonucleotides and gene editing |
| EP3215166B1 (en) | 2014-10-31 | 2024-04-24 | The Trustees of the University of Pennsylvania | Altering gene expression in car-t cells and uses thereof |
| WO2016097751A1 (en) | 2014-12-18 | 2016-06-23 | The University Of Bath | Method of cas9 mediated genome engineering |
| AU2016209295B2 (en) | 2015-01-21 | 2021-08-12 | Genevant Sciences Gmbh | Methods, compositions, and systems for delivering therapeutic and diagnostic agents into cells |
| EP3929296A1 (en) | 2015-01-30 | 2021-12-29 | The Regents of The University of California | Protein delivery in primary hematopoietic cells |
| US20180200387A1 (en) | 2015-02-23 | 2018-07-19 | Crispr Therapeutics Ag | Materials and methods for treatment of human genetic diseases including hemoglobinopathies |
| WO2016172359A2 (en) | 2015-04-24 | 2016-10-27 | The Regents Of The University Of California | Systems for detecting, monitoring or treating diseases or conditions using engineered cells and methods for making and using them |
| CA2986060A1 (en) | 2015-05-29 | 2016-12-08 | Valerie Odegard | Composition and methods for regulating inhibitory interactions in genetically engineered cells |
| AU2016278982A1 (en) | 2015-06-17 | 2018-01-18 | The Uab Research Foundation | CRISPR/Cas9 complex for genomic editing |
| EP3310931B1 (en) | 2015-06-17 | 2021-11-17 | The UAB Research Foundation | Crispr/cas9 complex for introducing a functional polypeptide into cells of blood cell lineage |
| US20170000743A1 (en) | 2015-07-02 | 2017-01-05 | Vindico NanoBio Technology Inc. | Compositions and Methods for Delivery of Gene Editing Tools Using Polymeric Vesicles |
| AU2016291778B2 (en) | 2015-07-13 | 2021-05-06 | Sangamo Therapeutics, Inc. | Delivery methods and compositions for nuclease-mediated genome engineering |
| US9512446B1 (en) | 2015-08-28 | 2016-12-06 | The General Hospital Corporation | Engineered CRISPR-Cas9 nucleases |
| US20180243446A1 (en) | 2015-09-01 | 2018-08-30 | The Hospital For Sick Children | Method and compositions for removing duplicated copy number variaions (cnvs) for genetic disorders and related uses |
| WO2017053729A1 (en) | 2015-09-25 | 2017-03-30 | The Board Of Trustees Of The Leland Stanford Junior University | Nuclease-mediated genome editing of primary cells and enrichment thereof |
| US11286480B2 (en) | 2015-09-28 | 2022-03-29 | North Carolina State University | Methods and compositions for sequence specific antimicrobials |
| EP4108255A1 (en) | 2015-10-05 | 2022-12-28 | Precision Biosciences, Inc. | Genetically-modified cells comprising a modified human t cell receptor alpha constant region gene |
| WO2017070169A1 (en) | 2015-10-19 | 2017-04-27 | The Methodist Hospital | Crispr-cas9 delivery to hard-to-transfect cells via membrane deformation |
| CN108368522A (en) | 2015-10-20 | 2018-08-03 | 10X 基因组学有限公司 | Method and system for high-throughput unicellular genetic manipulation |
| WO2017070429A1 (en) | 2015-10-22 | 2017-04-27 | Regents Of The University Of Minnesota | Methods involving editing polynucleotides that encode t cell receptor |
| CN108368520B (en) | 2015-11-04 | 2023-01-17 | 菲特治疗公司 | Genome engineering of pluripotent cells |
| JP2019500899A (en) | 2015-11-23 | 2019-01-17 | ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア | Cellular RNA tracking and manipulation through nuclear delivery of CRISPR / Cas9 |
| AU2016369490C1 (en) | 2015-12-18 | 2021-12-23 | Sangamo Therapeutics, Inc. | Targeted disruption of the T cell receptor |
| AU2016380836A1 (en) | 2015-12-30 | 2018-07-12 | Avectas Limited | Vector-free delivery of gene editing proteins and compositions to cells and tissues |
| US20190241910A1 (en) | 2016-03-11 | 2019-08-08 | Bluebird Bio, Inc. | Genome edited immune effector cells |
| JP2019510503A (en) | 2016-04-07 | 2019-04-18 | ブルーバード バイオ， インコーポレイテッド | Chimeric antigen receptor T cell composition |
| KR20230088514A (en) | 2016-04-14 | 2023-06-19 | 프레드 허친슨 캔서 센터 | Compositions and methods to program therapeutic cells using targeted nucleic acid nanocarriers |
| US10188749B2 (en) | 2016-04-14 | 2019-01-29 | Fred Hutchinson Cancer Research Center | Compositions and methods to program therapeutic cells using targeted nucleic acid nanocarriers |
| EP4180519A1 (en) | 2016-04-15 | 2023-05-17 | Memorial Sloan Kettering Cancer Center | Transgenic t cell and chimeric antigen receptor t cell compositions and related methods |
| WO2017189336A1 (en) | 2016-04-25 | 2017-11-02 | The Regents Of The University Of California | Methods and compositions for genomic editing |
| JP7148494B2 (en) * | 2016-04-25 | 2022-10-05 | ウニベルシテート バーゼル | Allele editing and its applications |
| WO2017186550A1 (en) | 2016-04-29 | 2017-11-02 | Basf Plant Science Company Gmbh | Improved methods for modification of target nucleic acids |
| WO2017210334A1 (en) | 2016-05-31 | 2017-12-07 | Massachusetts Institute Of Technology | Hydrodynamically controlled electric fields for high throughput transformation & high throughput parallel transformation platform |
| WO2017220527A1 (en) | 2016-06-20 | 2017-12-28 | Glycotope Gmbh | Means and methods for modifying multiple alleles |
| CN110312799A (en) | 2016-08-17 | 2019-10-08 | 博德研究所 | Novel C RISPR enzyme and system |
| US20190177677A1 (en) | 2016-08-20 | 2019-06-13 | The Regents Of The University Of California | High-throughput system and method for the temporary permeablization of cells |
| CN110291100A (en) | 2016-10-12 | 2019-09-27 | 费尔丹生物公司 | For polypeptide load to be delivered to the synthetic peptide shuttling agent of the rational design of the cytosol and/or nucleus of target eukaryocyte, purposes, relative method and kit from extracellular space |
| WO2018073393A2 (en) | 2016-10-19 | 2018-04-26 | Cellectis | Tal-effector nuclease (talen) -modified allogenic cells suitable for therapy |
| WO2018073391A1 (en) | 2016-10-19 | 2018-04-26 | Cellectis | Targeted gene insertion for improved immune cells therapy |
| WO2018094291A1 (en) | 2016-11-18 | 2018-05-24 | Christopher Bradley | Massively multiplexed homologous template repair for whole-genome replacement |
| US10975388B2 (en) | 2016-12-14 | 2021-04-13 | Ligandal, Inc. | Methods and compositions for nucleic acid and protein payload delivery |
| CN110546495B (en) | 2016-12-30 | 2022-11-01 | 加利福尼亚州立大学董事会 | Methods for selection and passage of genome editing T cells |
| WO2019084552A1 (en) | 2017-10-27 | 2019-05-02 | The Regents Of The University Of California | Targeted replacement of endogenous t cell receptors |
| BR112020008704A2 (en) | 2017-10-30 | 2020-11-10 | Pact Pharma, Inc. | editing primary cell genes |
-
2018
- 2018-06-15 WO PCT/US2018/037919 patent/WO2018232356A1/en unknown
- 2018-06-15 KR KR1020207001148A patent/KR20200018645A/en not_active Application Discontinuation
- 2018-06-15 CN CN201880053088.XA patent/CN111344020A/en active Pending
- 2018-06-15 CA CA3067382A patent/CA3067382A1/en active Pending
- 2018-06-15 EP EP18817563.2A patent/EP3638317A4/en active Pending
- 2018-06-15 SG SG11201912179SA patent/SG11201912179SA/en unknown
- 2018-06-15 US US16/622,843 patent/US20200362355A1/en not_active Abandoned
- 2018-06-15 AU AU2018283405A patent/AU2018283405A1/en active Pending
- 2018-06-15 KR KR1020247005507A patent/KR20240027153A/en active Application Filing
- 2018-06-15 JP JP2019569305A patent/JP7275054B2/en active Active
- 2018-06-15 MX MX2019015188A patent/MX2019015188A/en unknown
- 2018-06-15 BR BR112019026625-9A patent/BR112019026625A2/en unknown
-
2019
- 2019-12-12 IL IL271389A patent/IL271389A/en unknown
-
2021
- 2021-05-20 US US17/326,252 patent/US20210388362A1/en not_active Abandoned
- 2021-10-11 US US17/498,531 patent/US11814624B2/en active Active
-
2023
- 2023-05-02 JP JP2023076182A patent/JP2023100828A/en active Pending
- 2023-09-08 US US18/463,589 patent/US20240117360A1/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Gasser et al. | Expression of abbreviated mouse dihydrofolate reductase genes in cultured hamster cells. | |
| CN107446923B (en) | rAAV8-CRISPR-SaCas9 system and application thereof in preparation of hepatitis B treatment drug | |
| CN107893074A (en) | A kind of gRNA, expression vector, knockout system, kit for being used to knock out CXCR4 genes | |
| JP2016500262A5 (en) | ||
| WO2017075475A1 (en) | Crispr/cas-related methods and compositions for treating herpes simplex virus | |
| CA3001351A1 (en) | Crispr/cas-related methods and compositions for treating hepatitis b virus | |
| JP2018518182A5 (en) | ||
| Baur et al. | Intermolecular homologous recombination in plants | |
| CN107787367A (en) | Guiding RNA for the chemical modification of the gene regulation of CRISPR/CAS mediations | |
| WO2015153780A1 (en) | Crispr/cas-related methods and compositions for treating primary open angle glaucoma | |
| JP2020524998A5 (en) | ||
| BR112015022522B1 (en) | METHOD FOR MODIFICATION OF GENOMIC MATERIAL IN A PLANT CELL AND VIRAL VECTOR | |
| CA3025523A1 (en) | Peptides and nanoparticles for intracellular delivery of genome-editing molecules | |
| WO2015115903A1 (en) | Site-specific dna break-induced genome editing using engineered nucleases | |
| WO2012156839A2 (en) | New generation of splice-less lentiviral vectors for safer gene therapy applications | |
| JPWO2018232356A5 (en) | ||
| EP2872528A1 (en) | Compositions and methods for duchenne muscular dystrophy gene therapy | |
| Aksoy et al. | Multiple copies of a retroposon interrupt spliced leader RNA genes in the African trypanosome, Trypanosoma gambiense. | |
| CN111718964A (en) | Nucleic acid sequence and system for repairing DMD gene mutation | |
| CA3102072A1 (en) | Gene targeting | |
| CN110551762A (en) | CRISPR/ShaCas9 gene editing system and application thereof | |
| CN115820642A (en) | CRISPR-Cas9 system for treating duchenne muscular dystrophy | |
| Monstein et al. | A candidate gene for human U1 RNA. | |
| Khromov et al. | Delivery of CRISPR/Cas9 ribonucleoprotein complex to apical meristem cells for DNA-free editing of potato Solanum tuberosum genome | |
| CN107841509A (en) | A kind of kit for knocking out glioblastoma DHC2 genes |