JP2021511824A - 延長された単一ガイドrna及びその用途 - Google Patents
延長された単一ガイドrna及びその用途 Download PDFInfo
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Abstract
Description
1)前記細胞をデアミナーゼコーディングDNA、RNA−ガイドヌクレアーゼコーディングDNA、及び延長されたガイドRNAコーディング遺伝子をそれぞれ含んだり、或いはこれらのうち2つ以上を含む組換えベクターで形質感染させたり、
2)前記細胞にデアミナーゼ、RNA−ガイドヌクレアーゼ、及び延長されたガイドRNA(例えば、デアミナーゼ、RNA−ガイドヌクレアーゼ、及び延長されたガイドRNAを含む混合物又は複合体形態のリボ核酸タンパク質)を直接注入したり、或いは
3)前記細胞にデアミナーゼコーディングmRNA、RNA−ガイドヌクレアーゼコーディングmRNA及びガイドRNAの混合物又はこれらのそれぞれを直接注入して行うことができる。
(1)D10、H840、又はD10+H840;
(2)D1135、R1335、T1337、又はD1135+R1335+T1337;又は
(3)(1)及び(2)残基の両方、でアミノ酸置換が起こったものであり得る。
(1)D10又はH840位置に突然変異(例えば、他のアミノ酸への置換)が導入されてエンドヌクレアーゼ活性を喪失しニッカーゼ活性を有する変形Cas9、又はストレプトコッカスピオゲネス(Streptococcus pyogenes)由来のCas9タンパク質に、D10及びH840位置にともに突然変異(例えば、他のアミノ酸への置換)が導入されてエンドヌクレアーゼ活性及びニッカーゼ活性をともに喪失した変形Cas9タンパク質;
(2)D1135、R1335及びT1337の一つ以上又はこれら全部に突然変異(例えば、他のアミノ酸への置換)が導入されて野生型と異なるPAM配列を認識する変形Cas9タンパク質;又は
(3)(1)及び(2)の突然変異がともに導入されてニッカーゼ活性を有し、野生型と異なるPAM配列を認識するか、エンドヌクレアーゼ活性及びニッカーゼ活性をともに喪失し、野生型と異なるPAM配列を認識する変形Cas9タンパク質であり得る。
標的配列とハイブリダイズ可能な部位(標的化配列)を含むCRISPR RNA(crRNA);
Casタンパク質、Cpf1などのようなヌクレアーゼと相互作用する部位を含むトランス活性化(trans−activating)crRNA(tracrRNA);及び
前記crRNA及びtracrRNAの主要部位(例えば、標的化配列を含むcrRNA部位及びヌクレアーゼと相互作用するtracrRNAの部位)が融合された形態の単一ガイドRNA(single guide RNA;sgRNA)からなる群から選ばれる1種以上であり得、
具体的にCRISPR RNA(crRNA)及びトランス活性化(trans−activating)crRNA(tracrRNA)を含む二重RNA(dual RNA)、又はcrRNA及びtracrRNAの主要部位を含む単一ガイドRNA(sgRNA)であり得る。
一例において、前記ガイドRNAは次の一般式1で表現され得る:
5’−(Ncas9)l−(GUUUUAGAGCUA)−(オリゴヌクレオチドリンカー)−(UAGCAAGUUAAAAUAAGGCUAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGC(配列番号1)−3’ (一般式1)
(Ncas9)lは、Ncas9が標的遺伝子(target gene)の標的部位(target site)上に結合(ハイブリダイズ)する標的化配列であり、その核酸配列は前記標的部位の配列によって決定され(すなわち、標的部位の配列とハイブリダイズ可能な配列である。)、lが前記標的化配列に含まれたヌクレオチド数を示すものであって、20であり得、5’−末端から最初の核酸は標的部位配列とマッチングであるグアニン(Gで表示;標的部位の対応位置がシトシン(C)である場合)又はミスマッチングであるグアニン(gで表示;標的部位の対応位置がシトシン(C)でない場合)であり;
前記オリゴヌクレオチドリンカーは、3〜5個、例えば4個のヌクレオチドを含むものであり得、前記ヌクレオチドは互いに同一でも異なってもよく、A、U、C及びGからなる群からそれぞれ独立して選択され得る。
以下、実施例を用いて本発明をより詳細に説明する。これらの実施例は単に本発明をより具体的に説明するためのものであり、本発明の要旨によって本発明の範囲がこれらの実施例によって制限されないということは当業界における通常の知識を有する者にとって明らかであろう。
一例において、前記ガイドRNAは次の一般式1で表現され得る:
5’−(Ncas9)l−(GUUUUAGAGCUA)−(オリゴヌクレオチドリンカー)−(UAGCAAGUUAAAAUAAGGCUAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGC(配列番号60)−3’ (一般式1)
Claims (17)
- 標的配列とハイブリダイズ可能な延長されたガイドRNAであって、5’末端に1〜3個のグアニン(G)及び1〜10個のヌクレオチド(それぞれ独立してA、T、C、及びGの中から選択可能)をさらに含むことを特徴とする塩基校正用ガイドRNA。
- 前記5’末端に追加されるグアニンは標的配列と相補的であるか又は非相補的であることを特徴とする、請求項1に記載のガイドRNA。
- 前記ガイドRNAは、標的配列とハイブリダイズ可能な部位であるCRISPR RNA(crRNA)及び前記RNA−ガイドヌクレアーゼと相互作用する部位であるトランス活性化(trans−activating)crRNA(tracrRNA)を含むことを特徴とする、請求項1に記載のガイドRNA。
- (i)デアミナーゼ又はこれをコードする遺伝子、(ii)RNA−ガイドヌクレアーゼ又はこれをコードする遺伝子及び(iii)標的配列とハイブリダイズ可能な延長されたガイドRNA又はこれをコードする遺伝子を含む塩基校正用組成物であって、
前記延長されたガイドRNAは、5’末端に1〜3個のグアニン(G)及び1〜10個のヌクレオチド(それぞれ独立してA、T、C、及びGの中から選択可能)をさらに含むことを特徴とする塩基校正用組成物。 - 前記延長されたガイドRNAの5’末端に追加されるグアニンは、標的配列と相補的であるか又は非相補的であることを特徴とする、請求項4に記載の組成物。
- 前記ガイドRNAは、標的配列とハイブリダイズ可能な部位であるCRISPR RNA(crRNA)及び前記RNA−ガイドヌクレアーゼと相互作用する部位であるトランス活性化(trans−activating)crRNA(tracrRNA)を含むことを特徴とする、請求項4に記載の組成物。
- 前記ガイドRNAは、二重ガイドRNA又は単一ガイドRNA(sgRNA)であることを特徴とする、請求項4に記載の組成物。
- 前記デアミナーゼは、APOBEC1(apolipoprotein B editing complex 1)、AID(activation−induced deaminase)及びtadA(tRNA−specific adenosine deaminase)からなる群から選ばれることを特徴とする、請求項4に記載の組成物。
- ウラシルDNAグリコシラーゼ抑制剤(uracil DNA glycosylase inhibitor:UGI)又はこれをコードする遺伝子をさらに含むことを特徴とする、請求項4に記載の組成物。
- 核局在化配列(NLS)又はこれをコードする遺伝子をさらに含むことを特徴とする、請求項4に記載の組成物。
- 前記RNA−ガイドヌクレアーゼは、標的遺伝子の一方の鎖を切断するように変形された変形Cas9(CRISPR関連タンパク質9)又は変形Cpf1(プレボテラ(Prevotella)及びフランシセラ1(Francisella 1)由来CRISPR)システムであることを特徴とする、請求項4に記載の組成物。
- 前記RNA−ガイドヌクレアーゼは、Cas9ニッカーゼ(Cas9ニッカーゼ;nCas9)又は触媒活性欠乏Cas9(catalytically−deficient Cas9;dCas9)であることを特徴とする、請求項8に記載の組成物。
- 請求項4の塩基校正用組成物を細胞に導入させる段階を含む塩基校正方法。
- 前記細胞は真核細胞であることを特徴とする、請求項13に記載の方法。
- 前記真核細胞は動物細胞又は真核植物細胞であることを特徴とする、請求項14に記載の方法。
- 前記真核細胞は哺乳動物の胚細胞又は真核植物の胚であることを特徴とする、請求項15に記載の方法。
- 次の段階を含む突然変異が誘発されたヒト以外の哺乳動物又は真核植物の成体の作製方法;
(a)請求項4に記載の塩基校正用組成物を哺乳動物の胚又は真核植物の胚に導入させる段階;及び
(b)前記胚を成長させて成体を得る段階。
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Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2016500003A (ja) * | 2012-10-23 | 2016-01-07 | ツールゲン インコーポレイテッド | 標的dnaに特異的なガイドrnaおよびcasタンパク質コード核酸またはcasタンパク質を含む、標的dnaを切断するための組成物、ならびにその使用 |
WO2016134081A1 (en) * | 2015-02-18 | 2016-08-25 | Iowa State University Research Foundation, Inc. | Modification of transcriptional repressor binding site in nf-yc4 promoter for increased protein content and resistance to stress |
JP2016536021A (ja) * | 2013-11-07 | 2016-11-24 | エディタス・メディシン,インコーポレイテッド | CRISPR関連方法および支配gRNAのある組成物 |
JPWO2015133554A1 (ja) * | 2014-03-05 | 2017-04-06 | 国立大学法人神戸大学 | 標的化したdna配列の核酸塩基を特異的に変換するゲノム配列の改変方法及びそれに用いる分子複合体 |
WO2017070632A2 (en) * | 2015-10-23 | 2017-04-27 | President And Fellows Of Harvard College | Nucleobase editors and uses thereof |
JP2017520243A (ja) * | 2014-06-06 | 2017-07-27 | リジェネロン・ファーマシューティカルズ・インコーポレイテッドRegeneron Pharmaceuticals, Inc. | 標的遺伝子座を修飾するための方法及び組成物 |
JPWO2017183724A1 (ja) * | 2016-04-21 | 2019-02-28 | 国立大学法人神戸大学 | ゲノム配列改変技術における変異導入効率の向上方法、及びそれに用いる分子複合体 |
Family Cites Families (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008070663A2 (en) * | 2006-12-04 | 2008-06-12 | Abbott Laboratories | Companion diagnostic assays for cancer therapy |
US20100076057A1 (en) * | 2008-09-23 | 2010-03-25 | Northwestern University | TARGET DNA INTERFERENCE WITH crRNA |
US20150166982A1 (en) * | 2013-12-12 | 2015-06-18 | President And Fellows Of Harvard College | Methods for correcting pi3k point mutations |
CN105899657A (zh) * | 2013-12-12 | 2016-08-24 | 布罗德研究所有限公司 | 用于改变基因产物表达的crispr-cas系统和方法、结构信息以及诱导型模块化cas酶 |
JP2015133554A (ja) | 2014-01-10 | 2015-07-23 | 三菱電機株式会社 | 有線伝送装置及び終端抵抗の抵抗値の調整方法 |
KR101785847B1 (ko) * | 2015-05-12 | 2017-10-17 | 연세대학교 산학협력단 | 선형 이중가닥 DNA를 활용한 CRISPR/Cas9 시스템을 이용한 표적 유전체 교정 |
US9790490B2 (en) * | 2015-06-18 | 2017-10-17 | The Broad Institute Inc. | CRISPR enzymes and systems |
WO2017099494A1 (ko) * | 2015-12-08 | 2017-06-15 | 기초과학연구원 | Cpf1을 포함하는 유전체 교정용 조성물 및 그 용도 |
JP6826930B2 (ja) | 2016-03-29 | 2021-02-10 | 住友化学株式会社 | 発光素子 |
US11192929B2 (en) * | 2016-12-08 | 2021-12-07 | Regents Of The University Of Minnesota | Site-specific DNA base editing using modified APOBEC enzymes |
CN106834341B (zh) * | 2016-12-30 | 2020-06-16 | 中国农业大学 | 一种基因定点突变载体及其构建方法和应用 |
-
2019
- 2019-01-23 EP EP19743145.5A patent/EP3744844A4/en active Pending
- 2019-01-23 US US16/964,277 patent/US20210032621A1/en active Pending
- 2019-01-23 KR KR1020207021532A patent/KR20200103769A/ko not_active Application Discontinuation
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- 2019-01-23 JP JP2020561562A patent/JP7075170B2/ja active Active
- 2019-01-23 CN CN201980014548.2A patent/CN111742051A/zh active Pending
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2016500003A (ja) * | 2012-10-23 | 2016-01-07 | ツールゲン インコーポレイテッド | 標的dnaに特異的なガイドrnaおよびcasタンパク質コード核酸またはcasタンパク質を含む、標的dnaを切断するための組成物、ならびにその使用 |
JP2016536021A (ja) * | 2013-11-07 | 2016-11-24 | エディタス・メディシン,インコーポレイテッド | CRISPR関連方法および支配gRNAのある組成物 |
JPWO2015133554A1 (ja) * | 2014-03-05 | 2017-04-06 | 国立大学法人神戸大学 | 標的化したdna配列の核酸塩基を特異的に変換するゲノム配列の改変方法及びそれに用いる分子複合体 |
JP2017520243A (ja) * | 2014-06-06 | 2017-07-27 | リジェネロン・ファーマシューティカルズ・インコーポレイテッドRegeneron Pharmaceuticals, Inc. | 標的遺伝子座を修飾するための方法及び組成物 |
WO2016134081A1 (en) * | 2015-02-18 | 2016-08-25 | Iowa State University Research Foundation, Inc. | Modification of transcriptional repressor binding site in nf-yc4 promoter for increased protein content and resistance to stress |
WO2017070632A2 (en) * | 2015-10-23 | 2017-04-27 | President And Fellows Of Harvard College | Nucleobase editors and uses thereof |
JPWO2017183724A1 (ja) * | 2016-04-21 | 2019-02-28 | 国立大学法人神戸大学 | ゲノム配列改変技術における変異導入効率の向上方法、及びそれに用いる分子複合体 |
Non-Patent Citations (4)
Title |
---|
BANNO S. ET AL., NATURE MICROBIOLOGY, vol. 3, JPN6021037521, 5 February 2018 (2018-02-05), pages 423 - 429, ISSN: 0004601703 * |
KATO-INUI T. ET AL., NUCLEIC ACIDS RESEARCH, vol. 46, no. 9, JPN6021037525, 17 April 2018 (2018-04-17), pages 4677 - 4688, ISSN: 0004601704 * |
RAN F. ET AL., NATURE PROTOCOLS, vol. 8, no. 11, JPN6021037526, 2013, pages 2281 - 2308, ISSN: 0004601702 * |
RYU S. ET AL., NATURE BIOTECHNOLOGY, vol. 36, no. 6, JPN6021037523, 27 April 2018 (2018-04-27), pages 536 - 539, ISSN: 0004601705 * |
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KR20200103769A (ko) | 2020-09-02 |
JP7075170B2 (ja) | 2022-05-25 |
CN111742051A (zh) | 2020-10-02 |
US20210032621A1 (en) | 2021-02-04 |
WO2019147014A1 (ko) | 2019-08-01 |
EP3744844A1 (en) | 2020-12-02 |
EP3744844A4 (en) | 2021-10-20 |
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