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杩ワ涓界瀛╃ㄥ哄缂杈CRISPR/Cas9锛瀵规剁楠ㄩ跨哄锛MSTN锛杩琛浜查わ硅插轰袱杈剧澶у绁锛寤轰涓棣涓哄查ょ妯″

绉浜哄浣跨ㄧ哄缂杈锛椤惧涔锛藉璁╀汉绫诲哄杩琛缂杈锛瀹瀵圭瑰DNA娈电查ゃョCRISPR/Cas9浠ユワ灏辨跺哄缂杈姣浼匡硅磋璁や负藉ㄦ椿缁涓渚挎峰扳缂杈浠讳哄

涓涓濂╄杩CRISPR/Cas9

杩涓CRISPR/Cas9杩椤规涓娆″颁汉浠虫敞ㄦ涔锛璞璇哄涔绉扮2015骞村害界瀛村棰缁浜哄缁缂杈宸ュ封CRISPR/Cas9涓や缇コ绉瀛濡.寰峰炬肩航.澶般浜浜烘存寰浜2015骞村害瀛寮妗濂绱璇哄椋涔绉帮捐璁や负骞磋璐灏瀛绔浜

CRISPR/Cas9板椤逛涔涓轰藉寰濡姝よ锛浣浠ュㄧ袱涓骞存堕村锛灞涓虹╁棰绌跺伐蜂涓锛骞舵杩700绡稿宠琛瀹灏ュ浼濡浣褰卞版浠娲伙

CRISPR/Cas9户搁稿讹ZFN锛绫昏浆褰婵娲诲瀛搴╂搁搁讹TALEN锛涔虹绗浠ｂ哄缁瀹圭杈涓涓や唬告锛舵朵绠渚裤蹇烽浼癸璁╁杩椋′扮瀹楠瀹わ涓虹荤绛棰宸ュ枫

浜CRISPR/Cas绯荤垫ユ

CRISPR/Cas9垫ユ浜涓绉寰у郴缁涓轰钩ㄧ╃浜娆″绛绫讳技锛缁ㄦ垫姣澶婧璐ㄧヤ镜讹浼浜х稿璁板锛ユ垫璇ョ澶婧浼╄川娆″ヤ镜锛杩绉寰у辩CRISPR/Cas绯荤瀹

ㄧ哄缁涓锛瀛ㄧ涓茶撮澶搴锛杩浜澶搴稿淇瀹锛浠О涔涓CRISPR搴锛Clustered Regularly Interspersed Short Palindromic Repeats绨瑙寰撮澶搴锛

1.璁板ヤ镜妗ｆ

朵腑撮搴ユ浜姣澶婧璐ㄧ涓灏DNA锛瀵硅浜澶ュヤ镜璁板锛濡A绀猴

姣澶婧璐ㄧ涓锛瀛ㄢ撮搴锛撮搴姝ｆ浠稿搴撮搴骞朵虹锛杩浜撮搴涓ょ澶寤朵几涓⒈哄寰藉淇瀹锛浠О涓PAM锛Protospacer adjacent motifs-撮搴涓磋哄

褰姣澶婧璐ㄧDNA棣娆″ヤ镜扮浣讹缁浼瀵瑰婧DNA娼ㄧPAM搴杩琛灏涓磋PAM搴浣涓哄撮搴锛灏舵村扮哄缁涓CRISPR搴涓涓や釜澶搴涔淬杩灏辨撮搴浜х杩绋

2讳娆″ヤ镜

褰澶婧璐ㄧ姣娆″ヤ镜瀹夸富讹浼璇卞CRISPR搴琛ㄨ揪讹CRISPR搴杩杩涓缁淇瀹界哄锛绉颁负Cas哄CRISPR搴杞浜хCRISPR RNACas哄琛ㄨ揪浜х╃涓璧峰浣锛杩瀵PAM搴璇浠ュ撮搴涓婧DNA纰卞轰琛ラ瀵癸ユ惧板婧DNA涓跺锛骞跺跺诧瑙ｅ婧DNA杩涔灏卞浜瀵圭姣澶婧璐ㄧ娆″ヤ镜绛

姝ｆ轰杩绉ぉ插尽哄讹CRISPR/Cas9杩锛浠浣跨瀛浠╃RNA寮瀵Cas9搁搁跺瀵瑰绉缁哄缁瑰浣硅琛淇グ

涓CRISPR/Cas9瀹瑕浠涔锛

CRISPR/Cas9腑锛浠冲琚杈缁哄缁DNA浣姣澶婧DNA哄缂杈瀹瑕涓や釜宸ュ封瀵RNA锛guide RNA, gRNA锛Cas9姐

朵腑锛瀵RNA璁捐骞朵虹锛寰缂杈哄杩瑕瀛ㄧ稿淇瀹PAM搴锛充纰卞哄NGG锛朵腑N浠ユ换⒈猴锛涓瀵RNA瑕涓PAM涓娓哥搴纰卞轰琛ラ瀵广浠ュ哄查や负渚锛濡3绀猴ㄥ查ゅ哄涓涓娓稿璁捐涓″瀵RNA(瀵RNA1锛瀵RNA2)锛灏朵Cas9界哄璐ㄧ涓杞ョ涓瀵RNA杩纰卞轰琛ラ瀵瑰浠ラ跺PAM杩搴锛Cas9戒浣胯哄涓涓娓哥DNA炬

瀵逛DNA剧杩涓╀浠讹╀韩瀛ㄧDNA浼や慨澶搴绛哄讹浼灏涓涓娓镐袱绔搴杩璧锋ワ浠瀹浜缁涓哄查ゃ

DNAユ瀹圭瀹锛ㄦ虹涓涓虹渚涓涓慨澶妯℃胯川绮锛杩风灏变ф渚妯℃垮ㄤ慨澶杩绋涓ョ娈垫ユ瀹圭锛瀵瑰绮惧电杩琛哄缂杈锛骞跺跺ヤ唬瀛姣浣涓浠ュ哄缂杈ㄧ╂ā寤恒

褰讹CRISPR/Cas9骞堕惧涔俱瀵RNA跺搴瑰э浠ュDNA浼や慨澶涓纭э藉戒瀵艰村哄缁涓跺浣缃骇ョ绐锛涔灏辨璋遍垛璞★杩涔舵褰卞CRISPR/Cas9ㄧ堕涔涓浣绉浜哄涓Cas9界浼归锛瀵归跺哄璇瑰х澧寮猴 CRISPR/Cas9垛灏涓楂

CRISPR/Cas9浣锛

CRISPR/Cas9ㄥ荤ュ悍浜х娲汇瀹剁茬绛棰搴涓寰浜虹版

濡ㄥ荤ュ悍棰锛iPS缁锛璇卞澶藉共缁锛娌荤浜虹被板褰㈣传琛锛浠ュ浜虹ょ璇卞iPS缁锛╃CRISPR/Cas9瀵煎婧缁ヤ慨澶绐琛绾㈣藉哄锛灏淇iPS缁瀹璇卞涓洪琛骞茬绉绘扮浜轰姝ゅ锛娇CRISPR归HIV姣璇卞瀹缁姣寤虹妯″绛版Nature绛蹇ラㄥザ跺典腑锛╃CRISPR/Cas9澧寮哄佃浣插尽藉锛ㄥ茬归涔姝ｅㄥ╃ㄥ哄缂杈宸ュ烽杩瀵规捐褰卞瀹剁浜фц界哄浣硅琛硅浠ュ缇绛澶у瀹剁浜фц界楂绛

浣姝ｅ绉瀛锛浠讳版虹藉涓浜跺瀵硅瀛CRISPR/Cas9扮煎０讹涓灏浜轰瀵瑰轰璐ㄧ锛瑰惰遍朵浠跺藉村哄缁朵浣缃骇ョ琛ㄧず蹇с

浣涓虹界瀛涓椤归澶х达绗璁や负CRISPR/Cas9版сф搴哥疆锛瀵CRISPR绯荤璁よ娣憋瀹楠璁捐浼归锛镐俊舵舵浼杩涓姝ユ楂锛CRISPR/Cas9浠ュ惰绌朵甯ヤ虹瀛涓宸ㄥぇ┿寰ㄤ涔灏ワCRISPR/Cas9甯ョ宸ㄥぇ杞蹇灏藉娉戒瀹讹版跺灞浜璇哄缁灏版ャ

[1]Doudna, J.A.; Charpentier, E., Genome editing. The new frontier of genome engineering with CRISPR-Cas9. Science, 2014, 346, (6213), 1258096.[剧ユ][2]Fang, R.; Chang, F.; Sun, Z.-L.; Li, N.; Meng, Q.-Y., New Method of Genome Editing Derived From CRISPR/Cas9. Acta Agronomica Sinica, 2013, 40, (8), 691.

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