JP2002535995A5 - - Google Patents
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- JP2002535995A5 JP2002535995A5 JP2000597445A JP2000597445A JP2002535995A5 JP 2002535995 A5 JP2002535995 A5 JP 2002535995A5 JP 2000597445 A JP2000597445 A JP 2000597445A JP 2000597445 A JP2000597445 A JP 2000597445A JP 2002535995 A5 JP2002535995 A5 JP 2002535995A5
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- 229920003013 deoxyribonucleic acid Polymers 0.000 description 44
- 238000003776 cleavage reaction Methods 0.000 description 16
- 108091007521 restriction endonucleases Proteins 0.000 description 13
- 108091006028 chimera Proteins 0.000 description 11
- 230000004568 DNA-binding Effects 0.000 description 10
- 230000002759 chromosomal Effects 0.000 description 10
- 230000001939 inductive effect Effects 0.000 description 7
- 230000035772 mutation Effects 0.000 description 3
- 238000005215 recombination Methods 0.000 description 3
- 229920000160 (ribonucleotides)n+m Polymers 0.000 description 2
- 238000010240 RT-PCR analysis Methods 0.000 description 2
- 231100000722 genetic damage Toxicity 0.000 description 2
- 102220174584 rs2228570 Human genes 0.000 description 2
- HCHKCACWOHOZIP-UHFFFAOYSA-N zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 2
- 239000011701 zinc Substances 0.000 description 2
- 229910052725 zinc Inorganic materials 0.000 description 2
- BRZYSWJRSDMWLG-DJWUNRQOSA-N (2R,3R,4R,5R)-2-[(1S,2S,3R,4S,6R)-4,6-diamino-3-[(2S,3R,4R,5S,6R)-3-amino-4,5-dihydroxy-6-[(1R)-1-hydroxyethyl]oxan-2-yl]oxy-2-hydroxycyclohexyl]oxy-5-methyl-4-(methylamino)oxane-3,5-diol Chemical compound O1C[C@@](O)(C)[C@H](NC)[C@@H](O)[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H]([C@@H](C)O)O2)N)[C@@H](N)C[C@H]1N BRZYSWJRSDMWLG-DJWUNRQOSA-N 0.000 description 1
- 101700014779 GLB1 Proteins 0.000 description 1
- 229920000272 Oligonucleotide Polymers 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000000875 corresponding Effects 0.000 description 1
- 230000002068 genetic Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000006011 modification reaction Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
Description
【特許請求の範囲】
【請求項１】 （ａ）細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、目的の部位にて二本鎖切断を誘導する工程；および
（ｂ）目的の部位への標的ＤＮＡの導入に適切な条件下で細胞に標的ＤＮＡを導入する工程を含み、該標的ＤＮＡが（１）目的の部位の周囲の領域に相同なＤＮＡおよび（２）該標的ＤＮＡと染色体ＤＮＡとの間の組換えの際に目的の特定配列を修復するＤＮＡを含有する、
細胞の染色体ＤＮＡ中の目的の特定配列の修復方法。
【請求項２】 （ａ）細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、改変対象の特定配列中の目的の部位にて二本鎖切断を誘導する工程；および
（ｂ）目的の部位への標的ＤＮＡの導入に適した条件下で細胞に標的ＤＮＡを導入する工程を含み、該標的ＤＮＡが（１）目的の部位の周囲の領域に相同なＤＮＡおよび（２）該標的ＤＮＡと染色体ＤＮＡとの間の組換えの際に特定配列を改変するＤＮＡを含有する、
細胞の染色体ＤＮＡ中の特定配列の改変方法。
【請求項３】 （ａ）細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、目的の内因性遺伝子中の目的部位にて二本鎖切断を誘導する工程；および
（ｂ）目的の部位への標的ＤＮＡの導入に適した条件下で細胞に標的ＤＮＡを導入する工程を含み、該標的ＤＮＡが（１）目的の部位の周囲の領域に相同なＤＮＡおよび（２）該標的ＤＮＡと目的の遺伝子との間の組換えの際に目的の遺伝子を減弱化するＤＮＡを含有する、
細胞の目的の内因性遺伝子の減弱化方法。
【請求項４】 （ａ）細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、目的の部位にて二本鎖切断を誘導する工程；および
（ｂ）目的の部位への標的ＤＮＡの導入に適した条件下で細胞に標的ＤＮＡを導入する工程を含み、該標的ＤＮＡが（１）目的の部位の周囲の領域に相同なＤＮＡおよび（２）目的の部位に導入すべき変異を含有する、
細胞の染色体ＤＮＡ中の目的の部位への変異の導入方法。
【請求項５】 目的の部位の周囲の領域に相同な染色体ＤＮＡが目的の部位に導入されるために、および遺伝子損傷の補正のために適した条件下で、細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、遺伝子損傷の目的の部位にて二本鎖切断を誘導する工程を含む、細胞の染色体ＤＮＡ中の遺伝子損傷の補正方法。
【請求項６】 目的の部位の周囲の領域に相同な染色体ＤＮＡが目的の部位に導入されるために、および特定配列の改変のために適した条件下で、細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、改変対象の特定配列中の目的の部位にて二本鎖切断を誘導する工程を含む、細胞の染色体ＤＮＡ中の特定配列の改変方法。
【請求項７】 目的の部位の周辺の領域に相同な染色体ＤＮＡが目的の部位に導入されるために、および目的の特定配列の修復のために適した条件下で、細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、目的の部位にて二本鎖切断を誘導する工程を含む、細胞の染色体ＤＮＡ中の目的の特定配列の修復方法。
【請求項８】 目的の特定配列が変異である、請求項１または７記載の方法。
【請求項９】 ＤＮＡ結合配列が亜鉛フィンガードメインまたはメガヌクレアーゼ認識部位である、請求項１〜８いずれか記載の方法。
【請求項１０】 ＤＮＡ切断ドメインが制限エンドヌクレアーゼ切断ドメインである、請求項１〜９いずれか記載の方法。
【請求項１１】 メガヌクレアーゼ認識部位がＩ−ＳｃｅＩ認識部位であり、ＤＮＡ切断ドメインがＦｏｋＩ切断ドメインである、請求項９または１０記載の方法。
【請求項１２】 ＤＮＡ結合配列およびＤＮＡ切断ドメインを含有してなるキメラ制限エンドヌクレアーゼ。
【請求項１３】 ＤＮＡ結合配列が亜鉛フィンガードメインまたはメガヌクレアーゼ認識部位である、請求項１２記載のキメラ制限エンドヌクレアーゼ。
【請求項１４】 ＤＮＡ切断ドメインが制限エンドヌクレアーゼ切断ドメインである、請求項１２または１３記載のキメラ制限エンドヌクレアーゼ。
【請求項１５】 メガヌクレアーゼ認識部位がＩ−ＳｃｅＩ認識部位であり、ＤＮＡ切断ドメインがＦｏｋＩ切断ドメインである、請求項１３または１４記載のキメラ制限エンドヌクレアーゼ。
【請求項１】 （ａ）細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、目的の部位にて二本鎖切断を誘導する工程；および
（ｂ）目的の部位への標的ＤＮＡの導入に適切な条件下で細胞に標的ＤＮＡを導入する工程を含み、該標的ＤＮＡが（１）目的の部位の周囲の領域に相同なＤＮＡおよび（２）該標的ＤＮＡと染色体ＤＮＡとの間の組換えの際に目的の特定配列を修復するＤＮＡを含有する、
細胞の染色体ＤＮＡ中の目的の特定配列の修復方法。
【請求項２】 （ａ）細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、改変対象の特定配列中の目的の部位にて二本鎖切断を誘導する工程；および
（ｂ）目的の部位への標的ＤＮＡの導入に適した条件下で細胞に標的ＤＮＡを導入する工程を含み、該標的ＤＮＡが（１）目的の部位の周囲の領域に相同なＤＮＡおよび（２）該標的ＤＮＡと染色体ＤＮＡとの間の組換えの際に特定配列を改変するＤＮＡを含有する、
細胞の染色体ＤＮＡ中の特定配列の改変方法。
【請求項３】 （ａ）細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、目的の内因性遺伝子中の目的部位にて二本鎖切断を誘導する工程；および
（ｂ）目的の部位への標的ＤＮＡの導入に適した条件下で細胞に標的ＤＮＡを導入する工程を含み、該標的ＤＮＡが（１）目的の部位の周囲の領域に相同なＤＮＡおよび（２）該標的ＤＮＡと目的の遺伝子との間の組換えの際に目的の遺伝子を減弱化するＤＮＡを含有する、
細胞の目的の内因性遺伝子の減弱化方法。
【請求項４】 （ａ）細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、目的の部位にて二本鎖切断を誘導する工程；および
（ｂ）目的の部位への標的ＤＮＡの導入に適した条件下で細胞に標的ＤＮＡを導入する工程を含み、該標的ＤＮＡが（１）目的の部位の周囲の領域に相同なＤＮＡおよび（２）目的の部位に導入すべき変異を含有する、
細胞の染色体ＤＮＡ中の目的の部位への変異の導入方法。
【請求項５】 目的の部位の周囲の領域に相同な染色体ＤＮＡが目的の部位に導入されるために、および遺伝子損傷の補正のために適した条件下で、細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、遺伝子損傷の目的の部位にて二本鎖切断を誘導する工程を含む、細胞の染色体ＤＮＡ中の遺伝子損傷の補正方法。
【請求項６】 目的の部位の周囲の領域に相同な染色体ＤＮＡが目的の部位に導入されるために、および特定配列の改変のために適した条件下で、細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、改変対象の特定配列中の目的の部位にて二本鎖切断を誘導する工程を含む、細胞の染色体ＤＮＡ中の特定配列の改変方法。
【請求項７】 目的の部位の周辺の領域に相同な染色体ＤＮＡが目的の部位に導入されるために、および目的の特定配列の修復のために適した条件下で、細胞において、ＤＮＡ結合配列とＤＮＡ切断ドメインとを含むキメラ制限エンドヌクレアーゼを用いて、目的の部位にて二本鎖切断を誘導する工程を含む、細胞の染色体ＤＮＡ中の目的の特定配列の修復方法。
【請求項８】 目的の特定配列が変異である、請求項１または７記載の方法。
【請求項９】 ＤＮＡ結合配列が亜鉛フィンガードメインまたはメガヌクレアーゼ認識部位である、請求項１〜８いずれか記載の方法。
【請求項１０】 ＤＮＡ切断ドメインが制限エンドヌクレアーゼ切断ドメインである、請求項１〜９いずれか記載の方法。
【請求項１１】 メガヌクレアーゼ認識部位がＩ−ＳｃｅＩ認識部位であり、ＤＮＡ切断ドメインがＦｏｋＩ切断ドメインである、請求項９または１０記載の方法。
【請求項１２】 ＤＮＡ結合配列およびＤＮＡ切断ドメインを含有してなるキメラ制限エンドヌクレアーゼ。
【請求項１３】 ＤＮＡ結合配列が亜鉛フィンガードメインまたはメガヌクレアーゼ認識部位である、請求項１２記載のキメラ制限エンドヌクレアーゼ。
【請求項１４】 ＤＮＡ切断ドメインが制限エンドヌクレアーゼ切断ドメインである、請求項１２または１３記載のキメラ制限エンドヌクレアーゼ。
【請求項１５】 メガヌクレアーゼ認識部位がＩ−ＳｃｅＩ認識部位であり、ＤＮＡ切断ドメインがＦｏｋＩ切断ドメインである、請求項１３または１４記載のキメラ制限エンドヌクレアーゼ。
ｎｌｓｌａｃＺ遺伝子の存在に関して陽性である２４個のクローン中の１８個を突然変異ｎｌｓｌａｃＺ遺伝子の発現について評価した。突然変異ｎｌｓｌａｃＺ遺伝子の発現を評価するため、１８個の対応するジェネチシン耐性クローン培養物中の細胞からＲＮＡを抽出した。突然変異ｎｌｓｌａｃＺ遺伝子をコードするＲＮＡを逆転写酵素ポリメラー連鎖反応（ＲＴ−ＰＣＲ）で増幅した。オリゴヌクレオチドプライマー５’−ＴＡＣＡＣＧＣＧＴＣＧＴＧＡＴＴＡＧＣＧＣＣＧ−３’（配列番号：３）をｌａｃＺ逆転写に使用した。ハンレイおよびメルリー（Ｔ．Ｈａｎｌｅｙ ＆ Ｊ．Ｐ．Ｍｅｒｌｉｅ）（１９９１）、バイオフィードバック・イン・バイオテクニックス（ＢｉｏＦｅｅｄｂａｃｋ ｉｎ ＢｉｏＴｅｃｈｎｉｑｕｅｓ）、第１０巻、第１号、５６ページに記載通りにＰＣＲを行った。１８個のクローン中１１個で陽性反応を示した。
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US11866999P | 1999-02-03 | 1999-02-03 | |
| US60/118,669 | 1999-02-03 | ||
| PCT/US2000/003014 WO2000046386A2 (en) | 1999-02-03 | 2000-02-03 | Gene repair involving the induction of double-stranded dna cleavage at a chromosomal target site |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2002535995A JP2002535995A (ja) | 2002-10-29 |
| JP2002535995A5 true JP2002535995A5 (ja) | 2007-04-05 |
Family
ID=22380031
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2000597445A Pending JP2002535995A (ja) | 1999-02-03 | 2000-02-03 | 染色体標的部位での二本鎖ｄｎａ切断の誘導を含む遺伝子修復 |
Country Status (6)
| Country | Link |
|---|---|
| US (7) | US20020107214A1 (ja) |
| EP (1) | EP1147209A2 (ja) |
| JP (1) | JP2002535995A (ja) |
| AU (1) | AU2982900A (ja) |
| CA (1) | CA2361191A1 (ja) |
| WO (1) | WO2000046386A2 (ja) |
Families Citing this family (144)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2982900A (en) * | 1999-02-03 | 2000-08-25 | Children's Medical Center Corporation | Gene repair involving the induction of double-stranded dna cleavage at a chromosomal target site |
| WO2000046385A1 (en) | 1999-02-03 | 2000-08-10 | The Children's Medical Center Corporation | Gene repair involving in vivo excision of targeting dna |
| AU2003251286B2 (en) | 2002-01-23 | 2007-08-16 | The University Of Utah Research Foundation | Targeted chromosomal mutagenesis using zinc finger nucleases |
| US20060078552A1 (en) | 2002-03-15 | 2006-04-13 | Sylvain Arnould | Hybrid and single chain meganucleases and use thereof |
| WO2009095742A1 (en) | 2008-01-31 | 2009-08-06 | Cellectis | New i-crei derived single-chain meganuclease and uses thereof |
| US20030232410A1 (en) * | 2002-03-21 | 2003-12-18 | Monika Liljedahl | Methods and compositions for using zinc finger endonucleases to enhance homologous recombination |
| US7273758B2 (en) * | 2002-07-19 | 2007-09-25 | Centre National De La Recherche Scientifique | Methods for inducing targeted stimulation of meiotic recombination and kits for performing said methods |
| CA2497913C (en) | 2002-09-05 | 2014-06-03 | California Institute Of Technology | Use of chimeric nucleases to stimulate gene targeting |
| US7563600B2 (en) | 2002-09-12 | 2009-07-21 | Combimatrix Corporation | Microarray synthesis and assembly of gene-length polynucleotides |
| JP4966006B2 (ja) | 2003-01-28 | 2012-07-04 | セレクティス | カスタムメイドメガヌクレアーゼおよびその使用 |
| ITMI20030821A1 (it) * | 2003-04-18 | 2004-10-19 | Internat Ct For Genetic En Gineering And | Polipeptidi chimerici e loro uso. |
| US8409861B2 (en) | 2003-08-08 | 2013-04-02 | Sangamo Biosciences, Inc. | Targeted deletion of cellular DNA sequences |
| US20120196370A1 (en) | 2010-12-03 | 2012-08-02 | Fyodor Urnov | Methods and compositions for targeted genomic deletion |
| AU2007201649B2 (en) * | 2003-08-08 | 2010-11-04 | Sangamo Therapeutics, Inc. | Methods and Compositions for Targeted Cleavage and Recombination |
| US7888121B2 (en) | 2003-08-08 | 2011-02-15 | Sangamo Biosciences, Inc. | Methods and compositions for targeted cleavage and recombination |
| EP3222715A1 (en) * | 2003-08-08 | 2017-09-27 | Sangamo BioSciences, Inc. | Methods and compositions for targeted cleavage and recombination |
| US11311574B2 (en) | 2003-08-08 | 2022-04-26 | Sangamo Therapeutics, Inc. | Methods and compositions for targeted cleavage and recombination |
| SI2025756T1 (sl) | 2003-11-18 | 2011-10-28 | Bayer Bioscience Nv | Izboljšana ciljana vstavitev DNA v rastlinah |
| US7972854B2 (en) | 2004-02-05 | 2011-07-05 | Sangamo Biosciences, Inc. | Methods and compositions for targeted cleavage and recombination |
| WO2006074956A1 (en) * | 2005-01-14 | 2006-07-20 | Bayer Bioscience N.V. | Improved plant transformation methods |
| CN102286456B (zh) | 2005-04-04 | 2014-04-30 | 拜尔作物科学公司 | 用于去除所选择dna序列的方法和手段 |
| EP2484758B1 (en) | 2005-10-18 | 2013-10-02 | Precision Biosciences | Rationally-designed meganucleases with altered sequence specificity and DNA-binding affinity |
| WO2008027558A2 (en) | 2006-08-31 | 2008-03-06 | Codon Devices, Inc. | Iterative nucleic acid assembly using activation of vector-encoded traits |
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2000
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- 2000-02-03 JP JP2000597445A patent/JP2002535995A/ja active Pending
- 2000-02-03 CA CA002361191A patent/CA2361191A1/en not_active Abandoned
- 2000-02-03 EP EP00908499A patent/EP1147209A2/en not_active Ceased
- 2000-02-03 WO PCT/US2000/003014 patent/WO2000046386A2/en not_active Application Discontinuation
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2001
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2003
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2006
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-
2009
- 2009-10-28 US US12/607,502 patent/US20100111924A1/en not_active Abandoned
-
2012
- 2012-03-12 US US13/417,969 patent/US8921332B2/en not_active Expired - Fee Related
-
2014
- 2014-11-21 US US14/550,463 patent/US9458439B2/en not_active Expired - Fee Related
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2016
- 2016-09-30 US US15/281,242 patent/US20170037434A1/en not_active Abandoned
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