CA3046824A1 - Thermostable cas9 nucleases - Google Patents
Thermostable cas9 nucleases Download PDFInfo
- Publication number
- CA3046824A1 CA3046824A1 CA3046824A CA3046824A CA3046824A1 CA 3046824 A1 CA3046824 A1 CA 3046824A1 CA 3046824 A CA3046824 A CA 3046824A CA 3046824 A CA3046824 A CA 3046824A CA 3046824 A1 CA3046824 A1 CA 3046824A1
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- protein
- nucleic acid
- sequence
- cas protein
- target nucleic
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| WO2013066438A2 (en) | 2011-07-22 | 2013-05-10 | President And Fellows Of Harvard College | Evaluation and improvement of nuclease cleavage specificity |
| US9637739B2 (en) * | 2012-03-20 | 2017-05-02 | Vilnius University | RNA-directed DNA cleavage by the Cas9-crRNA complex |
| US20150044192A1 (en) | 2013-08-09 | 2015-02-12 | President And Fellows Of Harvard College | Methods for identifying a target site of a cas9 nuclease |
| US9359599B2 (en) | 2013-08-22 | 2016-06-07 | President And Fellows Of Harvard College | Engineered transcription activator-like effector (TALE) domains and uses thereof |
| US9388430B2 (en) | 2013-09-06 | 2016-07-12 | President And Fellows Of Harvard College | Cas9-recombinase fusion proteins and uses thereof |
| US9340799B2 (en) | 2013-09-06 | 2016-05-17 | President And Fellows Of Harvard College | MRNA-sensing switchable gRNAs |
| US9737604B2 (en) | 2013-09-06 | 2017-08-22 | President And Fellows Of Harvard College | Use of cationic lipids to deliver CAS9 |
| US11053481B2 (en) | 2013-12-12 | 2021-07-06 | President And Fellows Of Harvard College | Fusions of Cas9 domains and nucleic acid-editing domains |
| AU2015298571B2 (en) | 2014-07-30 | 2020-09-03 | President And Fellows Of Harvard College | Cas9 proteins including ligand-dependent inteins |
| EP3204496A1 (en) | 2014-10-10 | 2017-08-16 | Editas Medicine, Inc. | Compositions and methods for promoting homology directed repair |
| AU2015342749B2 (en) | 2014-11-07 | 2022-01-27 | Editas Medicine, Inc. | Methods for improving CRISPR/Cas-mediated genome-editing |
| EP3294896A1 (en) | 2015-05-11 | 2018-03-21 | Editas Medicine, Inc. | Optimized crispr/cas9 systems and methods for gene editing in stem cells |
| WO2016201047A1 (en) | 2015-06-09 | 2016-12-15 | Editas Medicine, Inc. | Crispr/cas-related methods and compositions for improving transplantation |
| GB201510296D0 (en) * | 2015-06-12 | 2015-07-29 | Univ Wageningen | Thermostable CAS9 nucleases |
| CA2999500A1 (en) | 2015-09-24 | 2017-03-30 | Editas Medicine, Inc. | Use of exonucleases to improve crispr/cas-mediated genome editing |
| US12043852B2 (en) | 2015-10-23 | 2024-07-23 | President And Fellows Of Harvard College | Evolved Cas9 proteins for gene editing |
| EP3433363A1 (en) | 2016-03-25 | 2019-01-30 | Editas Medicine, Inc. | Genome editing systems comprising repair-modulating enzyme molecules and methods of their use |
| EP4047092B1 (en) | 2016-04-13 | 2025-07-30 | Editas Medicine, Inc. | Cas9 fusion molecules, gene editing systems, and methods of use thereof |
| KR20250103795A (ko) | 2016-08-03 | 2025-07-07 | 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 | 아데노신 핵염기 편집제 및 그의 용도 |
| CN109804066A (zh) | 2016-08-09 | 2019-05-24 | 哈佛大学的校长及成员们 | 可编程cas9-重组酶融合蛋白及其用途 |
| US11542509B2 (en) | 2016-08-24 | 2023-01-03 | President And Fellows Of Harvard College | Incorporation of unnatural amino acids into proteins using base editing |
| AU2017342543B2 (en) | 2016-10-14 | 2024-06-27 | President And Fellows Of Harvard College | AAV delivery of nucleobase editors |
| CA3046824A1 (en) | 2016-12-14 | 2018-06-21 | Wageningen Universiteit | Thermostable cas9 nucleases |
| US12286727B2 (en) | 2016-12-19 | 2025-04-29 | Editas Medicine, Inc. | Assessing nuclease cleavage |
| WO2018119359A1 (en) | 2016-12-23 | 2018-06-28 | President And Fellows Of Harvard College | Editing of ccr5 receptor gene to protect against hiv infection |
| EP3565907B1 (en) | 2017-01-06 | 2022-05-04 | Editas Medicine, Inc. | Methods of assessing nuclease cleavage |
| EP3592853A1 (en) | 2017-03-09 | 2020-01-15 | President and Fellows of Harvard College | Suppression of pain by gene editing |
| EP3592381A1 (en) | 2017-03-09 | 2020-01-15 | President and Fellows of Harvard College | Cancer vaccine |
| KR20190127797A (ko) | 2017-03-10 | 2019-11-13 | 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 | 시토신에서 구아닌으로의 염기 편집제 |
| CA3057192A1 (en) | 2017-03-23 | 2018-09-27 | President And Fellows Of Harvard College | Nucleobase editors comprising nucleic acid programmable dna binding proteins |
| EP3615672A1 (en) | 2017-04-28 | 2020-03-04 | Editas Medicine, Inc. | Methods and systems for analyzing guide rna molecules |
| WO2018209320A1 (en) | 2017-05-12 | 2018-11-15 | President And Fellows Of Harvard College | Aptazyme-embedded guide rnas for use with crispr-cas9 in genome editing and transcriptional activation |
| CN110997908A (zh) | 2017-06-09 | 2020-04-10 | 爱迪塔斯医药公司 | 工程化的cas9核酸酶 |
| EP3652312A1 (en) | 2017-07-14 | 2020-05-20 | Editas Medicine, Inc. | Systems and methods for targeted integration and genome editing and detection thereof using integrated priming sites |
| CN111801345A (zh) | 2017-07-28 | 2020-10-20 | 哈佛大学的校长及成员们 | 使用噬菌体辅助连续进化(pace)的进化碱基编辑器的方法和组合物 |
| WO2019139645A2 (en) | 2017-08-30 | 2019-07-18 | President And Fellows Of Harvard College | High efficiency base editors comprising gam |
| GB201716590D0 (en) * | 2017-10-10 | 2017-11-22 | Univ Wageningen | Thermostable cas9 nucleases with reduced off-target activity |
| CA3082251A1 (en) | 2017-10-16 | 2019-04-25 | The Broad Institute, Inc. | Uses of adenosine base editors |
| EP3724214A4 (en) | 2017-12-15 | 2021-09-01 | The Broad Institute Inc. | SYSTEMS AND METHODS FOR PREDICTING REPAIR RESULTS IN GENETIC ENGINEERING |
| SG11202007382TA (en) * | 2018-02-15 | 2020-08-28 | Sigma Aldrich Co Llc | Engineered cas9 systems for eukaryotic genome modification |
| WO2019226953A1 (en) | 2018-05-23 | 2019-11-28 | The Broad Institute, Inc. | Base editors and uses thereof |
| CA3104856A1 (en) | 2018-06-29 | 2020-01-02 | Editas Medicine, Inc. | Synthetic guide molecules, compositions and methods relating thereto |
| US12281338B2 (en) | 2018-10-29 | 2025-04-22 | The Broad Institute, Inc. | Nucleobase editors comprising GeoCas9 and uses thereof |
| US20220010321A1 (en) | 2018-11-01 | 2022-01-13 | Keygene N.V. | Dual guide rna for crispr/cas genome editing in plants cells |
| AU2019380672A1 (en) * | 2018-11-16 | 2021-05-27 | Depixus | Optimization of In Vitro Isolation Of Nucleic Acids Using Site-Specific Nucleases |
| RU2712497C1 (ru) * | 2018-11-26 | 2020-01-29 | Автономная некоммерческая образовательная организация высшего образования Сколковский институт науки и технологий | Средство разрезания ДНК на основе Cas9 белка из биотехнологически значимой бактерии Clostridium cellulolyticum |
| US12351837B2 (en) | 2019-01-23 | 2025-07-08 | The Broad Institute, Inc. | Supernegatively charged proteins and uses thereof |
| BR112021016019A2 (pt) | 2019-02-15 | 2021-10-05 | Sigma-Aldrich Co. Llc | Sistemas e proteínas de fusão crispr/cas |
| DE112020001306T5 (de) | 2019-03-19 | 2022-01-27 | Massachusetts Institute Of Technology | Verfahren und zusammensetzungen zur editierung von nukleotidsequenzen |
| US12473543B2 (en) | 2019-04-17 | 2025-11-18 | The Broad Institute, Inc. | Adenine base editors with reduced off-target effects |
| CN116694603A (zh) * | 2019-05-14 | 2023-09-05 | 深圳华大生命科学研究院 | 新型的Cas蛋白、Crispr-Cas系统及其在基因编辑领域中的用途 |
| WO2021017200A1 (zh) * | 2019-07-30 | 2021-02-04 | 湖北大学 | 一种基于运动发酵单胞菌的CRISPR-Cas系统、基因组编辑体系及其应用 |
| CN110331158B (zh) * | 2019-07-30 | 2021-09-14 | 湖北大学 | 基于运动发酵单胞菌内源CRISPR-Cas系统的多基因位点同时编辑方法及其应用 |
| CN112410234B (zh) * | 2019-08-21 | 2022-08-23 | 江南大学 | 一种多靶点编辑重组曲霉菌株的可视化筛选方法 |
| US11879134B1 (en) * | 2019-09-05 | 2024-01-23 | The Regents Of The University Of Colorado, A Body Corporate | Recombineering machinery to increase homology directed genome editing in thermophilic microbes |
| US12435330B2 (en) | 2019-10-10 | 2025-10-07 | The Broad Institute, Inc. | Methods and compositions for prime editing RNA |
| JP2023525304A (ja) | 2020-05-08 | 2023-06-15 | ザ ブロード インスティテュート，インコーポレーテッド | 標的二本鎖ヌクレオチド配列の両鎖同時編集のための方法および組成物 |
| CN111778230A (zh) * | 2020-07-17 | 2020-10-16 | 山东舜丰生物科技有限公司 | 一种适用于Cas12蛋白的缓冲系统及其应用 |
| RU2749307C1 (ru) * | 2020-10-30 | 2021-06-08 | Федеральное государственное бюджетное научное учреждение "Всероссийский научно-исследовательский институт сельскохозяйственной биотехнологии" (ФГБНУ ВНИИСБ) | Новая компактная нуклеаза CAS9 II типа из Anoxybacillus flavithermus |
| CN113881652B (zh) * | 2020-11-11 | 2022-11-22 | 山东舜丰生物科技有限公司 | 新型Cas酶和系统以及应用 |
| CN114480347B (zh) * | 2022-02-21 | 2022-12-23 | 中国科学院地球化学研究所 | 一种纯化Cas12a蛋白的方法 |
| CN114934031B (zh) * | 2022-05-25 | 2023-08-01 | 广州瑞风生物科技有限公司 | 新型Cas效应蛋白、基因编辑系统及用途 |
| JP7152094B1 (ja) | 2022-06-30 | 2022-10-12 | リージョナルフィッシュ株式会社 | ｔｒａｃｒＲＮＡユニット、及びゲノム編集方法 |
| CN116083400B (zh) * | 2022-12-08 | 2023-12-12 | 广州瑞风生物科技有限公司 | Cas蛋白截短体、构建其的方法及其应用 |
| CN116144631B (zh) * | 2023-01-17 | 2023-09-15 | 华中农业大学 | 耐热型核酸内切酶及其介导的基因编辑系统 |
| WO2025107617A1 (zh) * | 2023-11-24 | 2025-05-30 | 华智生物技术有限公司 | CRISPR/Cas效应蛋白及系统 |
Family Cites Families (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2176082B1 (es) | 2000-05-31 | 2004-01-01 | Consejo Superior Investigacion | Bacterias con la capacidad de unir metales pesados y su empleo en la detoxificacion de medios contaminados con metales pesados. |
| US7220571B2 (en) | 2000-09-28 | 2007-05-22 | Archer-Daniels-Midland Company | Escherichia coli strains which over-produce L-threonine and processes for the production of L-threonine by fermentation |
| AU2002234513B2 (en) * | 2001-02-23 | 2006-12-07 | Novozymes A/S | Method of generating diversity into lipolytic enzymes and lipolytic enzyme genes |
| US7670807B2 (en) | 2004-03-10 | 2010-03-02 | East Tennessee State Univ. Research Foundation | RNA-dependent DNA polymerase from Geobacillus stearothermophilus |
| JP2006230303A (ja) | 2005-02-25 | 2006-09-07 | Art Engineering Kk | 好熱性リパーゼ産生菌およびその利用 |
| KR100872695B1 (ko) | 2006-11-27 | 2008-12-10 | 씨제이제일제당 (주) | Ｇｒａｓ 미생물로부터 발현된 식품안전형 호열성아라비노스 이성화효소 및 그를 이용한 타가토스의제조방법 |
| EP2468869B1 (en) * | 2007-01-31 | 2015-03-18 | Pfenex Inc. | Bacterial leader sequences for increased expression |
| IN2012DN02396A (enExample) | 2009-09-29 | 2015-08-21 | Butamax Tm Advanced Biofuels | |
| FI3597749T3 (fi) | 2012-05-25 | 2023-10-09 | Univ California | Menetelmiä ja koostumuksia rna-ohjattua kohde-dna-modifikaatiota varten ja rna-ohjattua transkription modulaatiota varten |
| ES2921207T3 (es) | 2013-03-15 | 2022-08-19 | Cibus Us Llc | Procedimientos y composiciones para aumentar la eficiencia de la modificación genética direccionada utilizando la reparación genética mediada por oligonucleótidos |
| CA2942407C (en) | 2014-03-14 | 2023-09-26 | Cibus Us Llc | Methods and compositions for increasing efficiency of targeted gene modification using oligonucleotide-mediated gene repair |
| US10725041B2 (en) | 2014-11-04 | 2020-07-28 | Versiti Blood Research Institute Foundation, Inc. | Method to bioengineer designer platelets using gene editing and stem cell methodologies |
| AU2015342749B2 (en) | 2014-11-07 | 2022-01-27 | Editas Medicine, Inc. | Methods for improving CRISPR/Cas-mediated genome-editing |
| ES2865268T3 (es) | 2014-12-17 | 2021-10-15 | Dupont Us Holding Llc | Composiciones y métodos para la edición eficaz de genes en E. coli utilizando sistemas de ARN guía/endonucleasa CAS en combinación con moldes de modificación de polinucleótido circulares |
| WO2016179038A1 (en) | 2015-05-01 | 2016-11-10 | Spark Therapeutics, Inc. | ADENO-ASSOCIATED VIRUS-MEDIATED CRISPR-Cas9 TREATMENT OF OCULAR DISEASE |
| EP3294877A1 (en) * | 2015-05-15 | 2018-03-21 | Pioneer Hi-Bred International, Inc. | Rapid characterization of cas endonuclease systems, pam sequences and guide rna elements |
| GB201510296D0 (en) | 2015-06-12 | 2015-07-29 | Univ Wageningen | Thermostable CAS9 nucleases |
| CA3046824A1 (en) | 2016-12-14 | 2018-06-21 | Wageningen Universiteit | Thermostable cas9 nucleases |
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| BR112019012165B1 (pt) | Uso de pelo menos uma molécula-guia de rna e uma proteína cas, método de ligação, clivagem, marcação ou modificação de um polinucleotídeo alvo de fita dupla, célula procariótica transformada, e, complexo de nucleoproteína |
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