QmҏWЂ炭`qσ}EX̖

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iwa {݁j
emailF<a href="mailto:ikawa@biken.osaka-u.ac.jp">ɐ쐳l</a>
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̈Zr[, <strong>3</strong>, e008 (2014)@<a href="http://dx.doi.org/10.7875/leading.author.3.e008">DOI: 10.7875/leading.author.3.e008</a>
Masahito Ikawa: <strong>Genome editing in mice opens a new era for biological and biomedical researches.</strong>

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<h2>v </h2>
@`qϓp̑햡́C|{זE⎎Ǔł݂͂Ȃ_Ci~bNȍۂC̃xŊώ@邢͉͂ł邱ƂɂDƂɁCqgƓMނɑ}EX́CSQmz񂪖炩ɂĂ邾łȂCESזEɂ鑊g𗘗pWI`q̔j󂪉\ł邱ƂC`q@\̓c[ƂĂłȂCqg̎̕aԃfƂāCȊwшw̑bSĂDɍŋ߂ł́CDPƂׂQmҏWZpCƂRNAɂWIzFCRISPR/Cas@̊JɂC1xF̃xł̈`q̑}CCuCɂ́CGtFN^[Ƃ̑gɂ]ʐCQm̉CGsQmC̑Ȃǂ̉\傫LĂD̃r[ł́C]̈`qσ}EX̍쐻Zpӂ܂CQmҏWЂ炭`qσ}EX̖ɂĉD

<h2>͂߂</h2>
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@̃xɂă}EX̃Qm`q𑀍삷鎎݂͌ÂC܂܂Ȏ@pĂDƂÓTIȕ@͎Rψك}EX̃XN[jOłCBk[h}EXCTזEBזESCID}EXȂǂLłDÂāCː̏Ǝ˂≻wψٌ̓^Ȃǂ̐lדˑRψٖ@JꂽDƂ΁CG`jg\AfiethylnitrosoureaFENUj̓^͎Rψق̖100{`1000{̊mœ_ψقUCkψقłȂA~m_uψقȂǂD@͊ȒPŁCG`jg\Af𓊗^YƖ쐶^̎zēq\^ɂXN[jODȂCQmɂĕψق𓯒肷邱Ƃ͔ɍłDߔN̉z͋Zp̔IȐiWɂƂȂC_ɓꂽψقɃXN[jOƂAv[`\ɂ͂ȂĂ邪<a href="#R1"><sup>1)</sup></a>Cvǂ̕ψق𓱓łȂƂ_͎cDȂCRψك}EXѐlדˑRψك}EX͂`qg̋KɂƂꂸC쐶^̃}EXƓlɈƂłD
@ψك}EXpĕ\^炻̌ƂȂ`qT肠Ă鏇`witH[hWFleBNXjɑ΂C`qςĕ\^Ƃ̊֘ATAv[`t`wio[XWFleBNXjƂD1970NɊÖ`q𓱓gXWFjbN}EXC1980N㔼ɓݐ̈`qj󂵂mbNAEg}EXJꂽƂɂCt`w͑傫iWD2010NɓCQmҏWZpp`qσ}EX̉\傫LĂDȉC`qg}EX̍쐻ZpƂ̉\ɂĉD

<h2>1DgXWFjbN}EX</h2>
@ʓIɂ́CgX`qƂ΂Ö`qh̃QmɐlדIɑgݍ݁CꂪĎ󂯌p}EXDh傪Ȃ^pN𔭌C쐶^邢͕ψٌ^̃^pNߏɔ肷ړIō쐻邱ƂDƂ΁CM҂炪Ju^pNEGFP𔭌O[}EX͈ڐA₪]ڂȂǕLbɁC܂CrasH2}EX͔񎎌ɗpĂDgDٓIȃv[^[ܗU^̃v[^[gƂɂC蕡GȈ`q̐\ƂȂD
@gXWFjbN}EX͎󐸗̑OjɃKXǂpĒDNA𒼐ڒ邱Ƃɂ쐻邱Ƃi<a href="#F1">}1a</a>jDgX`q̓Qm1ӏɃ^fɕ̃Rs[gݍ܂邱ƂC}ʒuRs[ɂ蔭p^[⋭xED͏ȂC`ECXxN^[󐸗Ɋ@gi<a href="#F1">}1b</a>jD̏ꍇ́CQm̌̕1Rs[gݍ܂邽߁Cʒuʂ𑊎E邱ƂłD܂COjɒ@ɔ׍쐻10{߂P邱ƂCHȕψك}EXngƂ1ł̔͂ȂǂɈЗ͂𔭊D

<a name="F1"></a><div id="fig1-caption-text" style="display: none;"><strong>}1@gXWFjbN}EX̍쐻@</strong>
iaj󐸗̑OjɃKXǂpĒDNA𒍓ƁC2َ̑eёٔՂɈ`qDQm1`2ӏɕ̃Rs[gݍ܂D
ibjт󐸗Ƀ`ECXxN^[ƁC7َ̑eёٔՂɈ`qDQm̉̕ӏ1Rs[gݍ܂D
icjтՖEɃ`ECXxN^[ƁCَeɈ`q𓱓邱ƂȂCقڂׂĂ̑ٔՂɈ`q𓱓łD
<a href="http://leading.lifesciencedb.jp/wordpress/wp-content/uploads/2014/07/Ikawa-3.e008-Fig.1.jpg" target="_blank">[Download]</a></div>[hs_figure id=1&amp;image=/wordpress/wp-content/uploads/2014/07/Ikawa-3.e008-Fig.1.png&amp;caption=fig1-caption-text]

@ȂC]̕@łَ͑eƑٔՂ̗Ɉ`q邽߁C󐫒vȂǂ̉͂łDM҂́C̐iՖEɃ`ECXxN^[΁CَeɈ`q邱ƂȂٔՂɓٓIɈ`q邱Ƃł邱Ƃoi<a href="#F1">}1c</a>jDDP̈`q@\̓c[ƂĊp҂D

<h2>2DmbNAEg}EX̊J</h2>
@mbNAEg}EX͓ݐ̈`qj󂳂ꂽ}EX̑̂ƂĎgĂC_ψقɂt[VtgCiZXψفC~XZXψقɂ@\̌ĈɏqׂQmҏWZpȂǂɂ`qj󂳂ꂽ}EX܂ށDʓIɂ́Cݐ̈`qj󂵂ESזEiembryonic stem cellC󐫊זEjL}EXĐBnɕψق`q`B}EXƂD
@1981NC}EXՖEESזECՖEɍ΃L}EX쐻ł邱ƁCɁCzɂESזEɗRq𓾂邱Ƃ񍐂ꂽD̂̂C܂ɂI邱Ƃ̂ł|{זE̗_𐶂CESזEp}EẌ`qj@ƂėL2̕@JꂽD_Ɉ`qj󂷂`qgbv@ƁCWIƂ`qj󂷂`q^[QeBO@łD`qgbv@́COܑ̖ϐ`qQmɃ_ɑ}ۂɁCRɓݐ̈`qɑ}ꂽ̂܂ɂID`q^[QeBO@́COܑ̖ϐ`q̗ɃQmƑ̈tCgɂݐ̈`qܑϐ`qƒu@łCړIƂ`q̋@\̃xŉ͂t`w̓T^ƂD1980N̏I肩1990NɂāCmbNAEg}EXɊւ鑽̘_gbvW[iɌfڂC̃xł̈`q̋@\͂ɂ̓mbNAEg}EX퓹Ƃ悤ɂȂD2007Nɂ́CmbNAEg}EX̊J҂łMartin EvansCOliver SmithiesCMario CapecchiɃm[xww܂^ĂD

<h2>3D`qgbv@</h2>
@`qgbv@ɂẮCܑ͖ϐ`qƃ|AtVOiȂJZbgpēݐ̃v[^[Ƃ炦v[^[gbv@嗬łCESזEŔĂ`qjłȂƂ_i<a href="#F2">}2a</a>jDōlĂꂽ̂CՓIɔv[^[̌Ƀ|[^[`q}JZbgpă|AƂ炦|Agbv@ŁCIɂׂ͂Ă̈`qgbvĔjłDC|Agbv@łܑ͖ϐ`q̂I~Rȟɓݐ̃GL\AȂ邽߁CiZXψوˑmRNA@\inon-sense mediated mRNA decayFNMDjɂmRNA̕s艻ЂNƂ肪D݂ł́Cܑϐ`q̌Ƀ{\[Gg[ʂ3t[ATGt邱ƂȂǂŖ͉Ă<a href="#R2"><sup>2)</sup></a>D`qgbvJZbg̓ɂ́CvX~h̃GNg|[V@p邪CזE̐⓱烌gECXxN^[g@D܂Dŋ߂́CSleeping BeautyPiggyBacȂǂ̃gX|]ngăQmɑgݍރAv[`JĂC̏ꍇ́C\^ώ@̂ɃgbvxN^[𔲂ċ@\̉񕜂mFłƂ_i<a href="#F2">}2a</a>jD

<a name="F2"></a><div id="fig2-caption-text" style="display: none;"><strong>}2@mbNAEg}EX̍쐻@</strong>
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ibj`q^[QeBO@DxN^[Ɠݐ̃QmƂ̂őgNƁCܑϐ`qJZbgWIƂȂ`qɑgݍ܂ȋIjDxN^[_ɑgݍ܂ƁCWteAőf邽ߍזE͎ʁȋIjD
icj`qgbvxN^[`q^[QeBOxN^[𓱓ESזE܂ɂIN[DPCR@TUubg@ɂCWI`qj󂳂ꂽESזEN[𓯒肵BD
idjF̂̐ł邱ƂmFESזEƋÏWCUDP}EXɈڐAăL}EX쐻D
iejꂽL}EXiF0j쐶^̃}EXƌzCF1ɂĕψو`q̓`BmFDɃweF1}EXǂzāCF2ɂăzmbNAEg}EX𓾂D
<a href="http://leading.lifesciencedb.jp/wordpress/wp-content/uploads/2014/07/Ikawa-3.e008-Fig.2.jpg" target="_blank">[Download]</a></div>[hs_figure id=2&amp;image=/wordpress/wp-content/uploads/2014/07/Ikawa-3.e008-Fig.2.png&amp;caption=fig2-caption-text]


<h2>4D`q^[QeBO@</h2>
@`qgbv@͓xN^[ԂESזEɓ邱Ƃɂ胉_Ɉ`qj󂷂̂ɑ΂C`q^[QeBO@ł͕WIƂ`qƂɃfUCꂽxN^[쐻ESזE֓Cܑϐ`qі܊󐫈`qi邢́CE`qjgݍ킹̑Iѕ̑Iɂ葊gNESזEZki<a href="#F2">}2b</a>jDɁC`q𓱓ESזE܂ɂIN[i<a href="#F2">}2c</a>jDN[ESזEQmDNA𒊏oPCR@TUubg@ɂ葊gmFDESזEƋÏWCUDP}EXɈڐAăL}EX쐻i<a href="#F2">}2d</a>jDCėpꂽ129nESזEC57BL/6nՖEɒăL}EX쐻ĂCC57BL/6nESזÉCICRn8זEƋÏWقBnɊ^₷悤łDꂽL}EX쐶^}EXƌzāCWI`qɕψقF1}EX𓾂̂CweF1}EX̎YzăzψقF2}EX𓾂i<a href="#F2">}2e</a>jDxN^[̍\zCESזEɂ鑊gCL}EX̍쐻ƌzɂ鐶Bnւ̊^CɁCzɂz}EX𓾂邽߂ɂ́CNPʂ̎ɂȂ邱ƂD
@}EXłESזE̔|{ZpC`qϋZpCL̍쐻ZpmĂ邪Cق̓ł͂ƂȂDƂ΃bgł́C}EXx邱15NC2003NɂȂĂ悤₭ESזEg`qmbNAEg񍐂ꂽDESזEȊO̍זEp`q^[QeBO@JĂDiPSזEpESזEƓl̎ł邱Ƃ͂܂łȂCGSזEigerm line stem cellCYBזEjɂĕWI`qj󂵁C𐸑ɈڐAĐq`𓾂邱Ƃł<a href="#R3"><sup>3)</sup></a>DȂCƒ{Ȃǂł́C`q^[QeBOَe@ۉזEN[Zpɂ̂𕜌@ʓIłD
@`qmbNAEg̑햡́C\^݂܂ŕWI`q̏dv킩ȂƂł낤DM҂܂Ő̃mbNAEg}EX쐻ĂC<em>in vitro</em>ɂ̌ʂNK{ƐMċ^Ȃ`qsvłC܂\ʌʂ琶ۂɕK{̈`q݂肵P[XȂȂDmbNAEg}EX̌ʂł͒ꔭt]ł邱ƂC\zǂ̕\^Cނ\zǑʂꂽق傫ȔɂȂD

<h2>5DۃmbNAEg}EXvWFNg</h2>
@`q^[QeBO̍ۂ̏dvȃ|CǵCBnɊ^ł関ȏԂێESזE̔|{ƁCIȑgłDESזE̔|{ɂẮCR}EX̌nꂽESזEȂǂɂĈقȂC|n⌌̃bgɂED̂߁C`qƌp̉ߒɂĐωăL}EXȂCL}EXaĂ`qςESזEɗReꂸCUoɂǂP[XȂȂDIȑgɂẮCǧグړIő̈𒷂Ƃ邽߁iv7`10 kbpjCS20 kbp߂vX~h̐؂\ƂȂCxN^[̍\zƔz̊mFɎԂƎԂvƂ_D܂Cg̕px͕WI`qxN^[̂̈̔zȂǂɂĂ傫قȂ邽߁CguV[eBO͌o⊨ɂ镔C^̂r̂킩ȂȂǏS҂ɂ͂ނłD
@ɎcOȂƂɁCRXgJ͂č쐻䂦ɁCꂽmbNAEg}EXޗƂċLȂCLȈ`q̃mbNAEg}EX͑̌ŏdč쐻ꂽD܂C\^݂̂ȂmbNAEg}EX͕̏\ɂłȂ߁CmbNAEg}EXقȂ錤łԂ쐻ƂނĂD
@̔wiCȂ_ɂĂׂĂ̈`qԗIɃmbNAEgĐlދʂ̍Yɂ悤ƂۃmbNAEg}EXvWFNgn<a href="#R4"><sup>4)</sup></a>Dč⃈[bp𒆐SɊLC2006NCIKMCiInternational Knockout Mouse ConsortiumjۃvWFNgƂĊ{iD͈`qgbv@嗬łCg̍xN^[̊JCxN^[\ž邢͋@BɂC`q^[QeBO@̗pĂDIKMCō̗pꂽ`q^[QeBOxN^[̑gDٓI邢͎ٓIɈ`qjłRfBVimbNAEg^CvłCݐ̃v[^[ɂ背|[^[Ƃ<em>LacZ</em>`q𔭌ƂӂȂĂD
@ȂC2011N獑ۃmbNAEg}EXvWFNgIMPCiInternational Mouse Phenotyping ConsortiumCURLF<a href="http://www.mousephenotype.org/" target="_blank">http://www.mousephenotype.org/</a>jɂЂpC܂łɁCIKMCƂ킹2߂`qɂđgESזEĂD́CL}EXmbNAEg}EX̍쐻1Iȕ\^̉͂̂ƂȂC͂̌ʂ͐EɌJD5N4000n̉͂IƂڕWݒ肳Ă邪CזExƂ͈قȂ̃x̎ɂ͔ȔpƎԂD2014N6_ł͖300n̕\^̌JɂƂǂ܂ĂC̐iW҂D

<h2>6DQmҏWZpƐlHyf</h2>
@Ċeȗ\Z𓊂čۃmbNAEg}EXvWFNg𐄐i邩CVȈ`qϋZpYĂDlHyf̊JƉpɂWI`q̉ϋZpC܂CQmҏWZpłDIɂ́CQm2{DNAؒfC̏C@\̃G[𗘗pĈ`qIɉςi<a href="#F3">}3a</a>jD񑊓[inonhomologous end-joiningFNHEJjoHɂCł́CؒfDNA[𒼐ڂȂ킹ۂɁČ邢͑}Ȃǂ̃G[N₷D܂Cgihomologous recombinationFHRjoHɂCł̓G[͏ȂC1{DNA邢2{DNAlדIɓĂƂ܂ăQmɎ荞܂邱ƂDQmҏWZpł́Ĉ悤2{DNAؒfƂ̏C̃G[܂pĈ`qςs<a href="#R5"><sup>5)</sup></a>D

<a name="F3"></a><div id="fig3-caption-text" style="display: none;"><strong>}3@QmҏWZp̌ƐlHyf</strong>
iajؒfʂ̂܂܌񑊓[oHɂCł͍Č̍ۂɌ}N₷DgoHɂC̍ۂɎQƂł悤1{DNA2{DNAƁC1̒umbNCȂǁCfUCψق𓱓邱ƂłD
ibjZFŃC3FWNtBK[`[t𐔌ƁCFokIkNA[[Z^pNƂč쐻D2ZFNZXуA`ZXFFokIʑ̂`CDNAؒf𔭊D
icjTALENł́CACCCGCTꂼ1ɓٓIɑΉ郆jbgȂTALGtFN^[WIƂȂDNAFD
idjCRISPR/Casnł́CgRNA5f[20WIDNAFDgRNAɃN[gꂽCas9́CPAMz̏㗬3Ԗڂ4Ԗڂ̉̂ؒfD
<a href="http://leading.lifesciencedb.jp/wordpress/wp-content/uploads/2014/07/Ikawa-3.e008-Fig.3.jpg" target="_blank">[Download]</a></div>[hs_figure id=3&amp;image=/wordpress/wp-content/uploads/2014/07/Ikawa-3.e008-Fig.3.png&amp;caption=fig3-caption-text]

@QmҏWƂZFNizinc finger nucleasejŏƎvlȂCȑOC2{DNAؒf𗘗p`qς͎݂ĂD1996Nɂ́C18ٓIɔFI-SceIKkNA[[p2{DNAؒfɂ葊ǧ̉P邱Ƃ񍐂ĂDCKkNA[[͕WIzfUC邱ƂłȂ߁CFzɃQmɑgݍޕKvȂǁCpIɂ͂قƂǈӖ̂Ȃ̂łD1996NɊJꂽ̂ZFNłDWNtBK[`[tƂ΂DNAhCFokIkNA[[ō\ꂽlHL^pNłCWI`qFؒfi<a href="#F3">}3b</a>jDCZFNłDNAhCfUCǂDNAFȂƂCxN^[̍\zނƂ̌_D܂Cs̕iɍzł߁Cʂ̌҂ɂ͍̉Ԃł邱ƂD
@ɓoꂵ̂CZFNDNAhCAa׋ۂł<em>Xanthomonas</em>׋ۂ̂TALEGtFN^[ɑウTALENłi<a href="#F3">}3c</a>jDj_ACCCGCTꂼ1ɓٓIɑΉ郆jbgWIzɂ킹đgݍ킹邾ō\zł邱ƂCʂ̌҂ɂpł郌xɂȂDɐF郂W[gݍ킹PȂĂD܂C34A~m_cȂ郆jbg4Ԗڂ32Ԗڂ̃A~m_c̃oG[VɒӂăW[\z邱ƂɂC̍TALEN悭쐻łPlatinum TALENJĂ<a href="#R6"><sup>6)</sup></a>iURLF<a href="http://www.addgene.org/TALEN/PlatinumGate/" target="_blank">http://www.addgene.org/TALEN/PlatinumGate/</a>jD
@ZFNTALENƂlHyf́C󐸗ɒ邾œݐ̈`qjł邱ƂCzCJGC[utBbVȂǁC̃fɂėpĂDڍׂɂĂ͖ɂQl}QƂꂽD

<h2>7DCRISPR/Cas@ɂQmҏWZp</h2>
@ZFNTALEN̓oɂglHyfhgQmҏWhƂt҂̎ڂ߁C܂CIȈ`qϋZpƂĂ̕]C2011Nɂ<em>Nature Method</em>Method of the YearC2012Nɂ<em>Science</em>Breakthrough of the yearɂƂ肠ꂽD̂悤ȏ󋵂̂ȂC̃QmҏWZpƂĊJꂽ̂CRISPR/Cas@łDCCRISPR/Casn͌Í׋ۂȂǂlƉu@\łCt@[WȂǂĐNODNAfЉĎ̃Qmɑgݍ݁C2xڂ̊̂ƂɁC̊ODNA̒fЂ甭2̏qRNApĕWIDNAFCCas9kNA[[N[gĂؒfD2013N2ɂ́C2̏qRNAЂƂ̃LRNAƂiguide RNAFgRNAC邢́Csingle guide RNAFsgRNAƂ΂jCCas9Ƃ2̈qŃQmDNA̐ؒf𔭊ł邱Ƃꂽ<a href="#R7"><sup>7,8)</sup></a>i<a href="#F3">}3d</a>jD
@ZFNTALENDNAhCƐؒfhCZ^pNƂċ@\邽ߕWI`qƂɕGȃxN^[̍\zKvȂ̂ɑ΂CCRISPR/Cas@ł͕WIzɑΉgRNA5f[20ύX邾ŕWI`qς邱ƂłD̎yƒRXgCؒfƍČCCRISPR/Cas@͏uԂɐȊw҂ɍL܂DM҂̓vX~hpX330iURLF<a href="http://www.addgene.org/42230/" target="_blank">http://www.addgene.org/42230/</a>jpĂ邪CpX330ɂgRNAJZbgуRhqgCas9JZbgڂĂCISDNA𓱓邾ŃmbNAEgxN^[\złi<a href="#F4">}4a</a>jD

<a name="F4"></a><div id="fig4-caption-text" style="display: none;"><strong>}4@CRISPR/Cas@ɂmbNAEg}EX̐vȍ쐻@</strong>
iajpX330vX~h́CqgU6v[^[̐䉺gRNACՐgAN`nCubhv[^[iCBhj̐䉺ɃRhqgCas9𔭌DWIƂPAMz̏㗬20<em>Bbs</em>Iyfʂɑ}邾ŃmbNAEgxN^[DpCAG-EGxxFP|[^[vX~hpX330vX~hƓɍזE֓ƁCpX330甭Cas9-gRNÂɂ背|[^[vX~hɑ}ꂽWIz񂪐ؒfDWIz̑Oɂd̈𗘗pCɂEGFPJZbgƁCזE͌u𔭂D
ibj󐸗̍זEgRNACas9mRNA𒍓RNA@ʓIł邪CpX330vX~h̂܂ܒ@ȕ֖@ƂĂĂDESזEP[XƂ͈قȂC󐸗̗̑Η`qɕψقꂽꍇɂF0ɂăzmbNAEg}EXȋΗ`q̕ψقƂ͂ȂjDȂĈ̂DNAؒfꂽꍇɂ́CUCN}EXƂĐ܂D
<a href="http://leading.lifesciencedb.jp/wordpress/wp-content/uploads/2014/07/Ikawa-3.e008-Fig.4.jpg" target="_blank">[Download]</a></div>[hs_figure id=4&amp;image=/wordpress/wp-content/uploads/2014/07/Ikawa-3.e008-Fig.4.png&amp;caption=fig4-caption-text]

@CRISPR/Cas@Mޔ|{זEɂċ@\邱ƂĂ甼NoȂ2013N5CCas9R[hmRNAgRNA󐸗̍זEɒ邱ƂɂmbNAEg}EXƂ_񍐂ꂽ<a href="#R9"><sup>9)</sup></a>i<a href="#F4">}4b</a>jDCM҂CRNÃXebvXLbvpX330vX~h󐸗̑OjɃ}CNCWFNVȈՖ@ƎɊJĂC2Ԏ̘_ƂƂŗ\zȏɐRC_\܂łɔNv<a href="#R10"><sup>10)</sup></a>DCRISPR/Cas@pmbNAEg}EX쐻̃bg͂̊ȕ֐ƃXs[hłCۂɁCISDNA̔pX330xN^[̍\zC|{זEɂؒf̌ɖ1TԁCɎ󐸗ւ̒oY܂ł3TԂ킦1JŃmbNAEg}EX쐻łDɁCψك}EX̖񔼐ɂė̑Η`qɕψقĂ_͓MׂłCESזEgƌzłNv邱ƂlƁCt@E_[Ńz}EX͖̂̂悤łDM҂̌oł́C100̎󐸗Ζ10C܂C5Cxψك}EXC2Cx̑Η`qɕψق<a href="#R10"><sup>10,11)</sup></a>DȂC󐸗ł͂ȂCESזEւ̃vX~h̃gXtFNVł́CpX330ߐɓꂽESזE̖8ŗ̑Η`qɕψقF߂D
@CRISPR/Cas@̒ӓ_ƂẮCWIƂzɋߎziIt^[QbgjؒfĂ܂XNDZFNTALENł́CZXƃA`ZXF2̃jbgɔFĂ͂߂FokIɂؒfN̂ɑ΂CБDNÂ݂FCRISPR/Cas@ِ̓͒ႢƂCזEȂǂ̔|{זEȂǂł̓It^[Qbg̐ؒf񍐂ĂDC󐸗ESזEȂǂłDNAC\͂̂CقǖɂȂȂ悤łDM҂ɂ󐸗ł̃mbNAEg}EX̍쐻ł́CIt^[QbǧƂ382ӏ𒲂ׂ3ӏ̐ؒfł<a href="#R11"><sup>11)</sup></a>D}EXł΁CzɂIt^[QbgɕψقȂ}EXIʂ邱Ƃ\ł邵CvɂΉłD邢́C`qWIƂ邪قȂgRNApĒǎ΁CIt^[Qbg̐ؒfɂe͖łł낤D

<h2>8DQmҏWZp̖</h2>
@2{DNAؒfDNAC̃G[𗘗pQmҏWZp̃bg͈`qjɂȂDؒfʂI[o[bv1{ISDNÂȂقǂɕψق𓱓Ă΁CWIψقݐ̃Qmɓ邱ƂłDCPȂDNAؒfɔׂĕψق̓͒ႢCESזEg킸Ɏ󐸗1ŕWIψق}EX쐻ł͖̂͂łD{Iɂ́C50قǂ̑100`130̃ISDNAgƂɂC1̒uFLAG^Ȏ}łi<a href="#F3">}3a</a>jD`qj󂷂ƒvƂȂ悤ȏꍇł_ψق̓Ȃ琶邱Ƃ\ɍlCqg̎Ƃ܂ψق𓱓邱ƂȂǂŎf}EXƂẲ\LĂDł́Czt]āC_ψق̈`qÂɎgꂽЉDt}AZg|_qh[[R[h`qɕψق߃`V𕪉ł̏QN`Vǃf}EXɁC̕ψٕʂؒfCas9gRNA𔭌vX~hƂƂɁC쐶^̔z1{DNAÖ˂ƂCꕔ̊̍זEɂĈ`qψقɂǂCɁCُȍזEƏXɒu邱Ƃɂ蓊򎡗Â̕Kv̂ȂȂƂ񍐂ꂽ<a href="#R12"><sup>12)</sup></a>D
@\悤ȒzWI`qɓꍇɂ́C1{DNAł͂ȂC2{DNA𓯎ɒ邱ƂɂȂi<a href="#F3">}3a</a>jDC2{DNAؒfɂ葊gPƂ͂C󐸗ւ2{DNA̒ɂ葊g}EX쐻邱Ƃ͂ނD悻20̏ɂă|[^[`q̔mFꂽƂ̕񍐂邪<a href="#R13"><sup>13)</sup></a>CM҂̌o⎄Mł́Cۂɐ܂Ă鑊g}EX͑ĂYe1`5ƍlDg}EXmɓɂ1000߂󐸗100C߂Y܂KvCRXg⓮̊ϓ_CȂ̉PƂ߂ł낤D_ł́CESזEɂ`q^[QeBÔƂ2{DNAؒfUČ}قpIȂD
@ƂŁC쐶^Cas9kNA[[2ӏ̍yfʂC2{DNA̗̍ʁXɐؒfDŁCЕ̊ʂj󂵂Cas9 nickasei<a href="#F5">}5a</a>jCсC̊ʂj󂵂dead Cas9쐻Ăi<a href="#F5">}5b</a>jDƂ΁CCas9 nickase2gRNAƑgݍ킹CꂼꂪZXуA`ZX̗ؒfƂɂ͂߂Ĉ`qjNƂɂCIt^[Qbg̐ؒfyƂAv[`񍐂Ă<a href="#R14"><sup>14)</sup></a>D܂Cdead Cas9WIƂȂ`qɌ邱Ƃɂ]ʐ̑jQ˂炤CRISPRi@<a href="#R15"><sup>15)</sup></a>i<a href="#F5">}5c</a>jCdead Cas9ɏCyfȂŃGsQmCς鎎<a href="#R16"><sup>16)</sup></a>i<a href="#F5">}5d</a>jC邢́Cdead Cas9GFPȂŕWI`q鎎݂񍐂Ă<a href="#R17"><sup>17)</sup></a>i<a href="#F5">}5e</a>jDZFNTALEgĕ񍐂ĂAv[`ł͂邪CgRNAς邾ł悢CRISPR/CasnƑgݍ킹邱ƂŔėp͔IɉPD܂זExł̎߁ĆC̃xł̉p҂D܂C2014NɂȂCCas9kNA[[̗̍\񍐂<a href="#R18"><sup>18)</sup></a>Cʂ̃Av[`\ƂȂƂCِ̉Pɂ킦ĐVȗp@̊J͂܂ĂD

<a name="F5"></a><div id="fig5-caption-text" style="display: none;"><strong>}5@QmҏW̉p</strong>
iajCas9 nickaseD2ӏ̍yfʂ̂Еj󂵂Cas9 nickase2gRNAƑgݍ킹ƁCZFNTALEN̂悤ɕWIِ̍ؒf\ɂȂD
ibjdead Cas9D2ӏ̍yfʂ𗼕Ƃj󂵂dead Cas9́CWIzƌ邪ؒf邱Ƃ͂łȂD
icj]ʐDdead Cas9WI`q̃v[^[Ɍ邱ƂɂC]ʂjQCRISPRi@񍐂ĂD
idjGsQm̏CDdead Cas9ɃGtFN^[Z邱ƂɂCWI`qDNA`邢̓qXg̏Cω邱ƂłD
iejC[WODdead Cas9EGFPZ邱ƂɂCWI`q邱ƂłD
ifjF̃xł̈`qρD2gRNACas9ɂC̈Ctʂ邢͓]NȂǁCF̃xł̈`qςsD
<a href="http://leading.lifesciencedb.jp/wordpress/wp-content/uploads/2014/07/Ikawa-3.e008-Fig.5.jpg" target="_blank">[Download]</a></div>[hs_figure id=5&amp;image=/wordpress/wp-content/uploads/2014/07/Ikawa-3.e008-Fig.5.png&amp;caption=fig5-caption-text]


<h2>9D̂ق̋Zp</h2>
@܂ł́CP̈`qɑ΂`qϋZpɂďЉCƃXP[̑傫Ȉ`qϋZpD܂Œ̈ꍇɂ́C̗̈ɂ܂`q^[QeBOxN^[pđgNƂD邢́CF̂2ӏ<em>loxP</em>ʂ}CCreRri[[ɂ肱𔲂̂ʓIłDꂪCRISPR/Cas@ł΁CQm2ӏɑ΂gRNAfUCCas9kNA[[ɂؒf邱ƂłȒPɎłi<a href="#F5">}5f</a>jDM҂60 kbp̗̈Ă邪CMłMbp̃xŌP[XD̏ꍇC͂N[5`10ŌƂp\ȌłD܂C̐F̂2ӏؒfƈ̕pxŋtʂ𓾂邱Ƃł邵CقȂF̂łΓ]U邱Ƃłi<a href="#F5">}5f</a>jD́C͂F̍Hẅɓł낤D
@F̍HwƂ΁ClHF̂𐶐Bnɓ`B邱Ƃ̂łgXN\~bN}EXJĂ<a href="#R19"><sup>19)</sup></a>DF̂ɂăNX^[\悤Ȗ򕨑ӊ֘AyfCqgR̈̐YȂǂɎgZpƂĒڂĂ邪Cǃ}EX̂̈`qςłDQmҏWZpƂ̑gɂiW҂D
@ƂŁC2{̍זEł͕ѕ̗̑Η`qɂĈ`qςKv邪C1{̍זEł1`q̔j󂠂邢͉ςɂ\^ώ@łD2011NC}EXɂĒPה󂩂1{̂ESזEC2{̉Α̍זE␶BזEɕ邱Ƃ̂ł邱Ƃꂽ<a href="#R20"><sup>20)</sup></a>DqgłKBM7זEHAP1זÊ悤1{̂ɋ߂ԂňێłזEmĂCQmҏWZpɂK͂Ȉ`qj󂪎݂ĂD1{̂ESזEIɈێł悤ɂȂΊpLł낤D

<h2></h2>
@MލזEɂCRISPR/Cas@@\邱Ƃ񍐂Ă1Nقǂ̂ɁCQmҏWZp͈`qσ}EX̏펯CȊwшw̌ɑ傫ȃCpNgD\Zʂ{iIȊɂ͂ĂȂ̂́C킪łCCRISPR/Cas@p̈`qϓɂiIȈw𐄐ivWFNgٓĂiURLF<a href="http://fmena.biken.osaka-u.ac.jp/" target="_blank">http://fmena.biken.osaka-u.ac.jp/</a>jDłɁCbgETMCƒ{CTłQmҏWZp̐񍐂ĂCf̊Jɂ킦Ö@ւ̉pȂǁC͑OՏ̐iɂՏ邢͑nւ̋n҂D

<h2> </h2>
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<h2>Ql}</h2>
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</ol>
<div class="au-profile">
<h2>҃vtB[</h2>
<span class="author">ɐ liMasahito Ikawaj</span>
F1997N ww@wȔmے CC2000N čThe Salk InstitutemC2004N wa oāC2011N蓯 D
FV`qϋZpJщpȂC󐸂𒆐SƂMނ̐B̋@\𖾂炩ɂD
URLF<a href="http://www.egr.biken.osaka-u.ac.jp/" target="_blank">http://www.egr.biken.osaka-u.ac.jp/</a>
</div>
? 2014 ɐ l Licensed under <a href="http://creativecommons.org/licenses/by/2.1/jp/" target="_blank">CC \ 2.1 {</a>
