24锛锛哄缂杈浣瑕ヤ锛

哄锛绉腑

朵锛哄缁澶绫宠哄绉 绠℃妤

讹涓界瀛㈣绠虹缁淇℃腑蹇

瑕涔扮锛剧ヨ缁锛

匡杩澶ヤ锛杩娌℃圭锛杩风瀵硅澶ф浠ユ案杩娑澶卞ㄨ甯轰

23ワ涓界瀛瀹ｅ锛浠浠杩颁浠ｅ哄缂杈宸ュCRISPR锛ヤ绉瑙ｅ惰1锛UCP1锛哄锛绉澧锛硅插轰瑰ュ悍姣姝ｅ父24锛

淇璇璇存浠ラ涓哄ぉ锛绉瀛涓浠村婊＄锛ｄ姝ゆ峰硅插烘ョ杩灏辫渚CRISPR/Cas9В充

CRISPR/Cas9

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

舵朵绠渚裤蹇烽浼癸璁╁杩椋′扮瀹楠瀹わ涓虹荤绛棰宸ュ凤磋璁や负藉ㄦ椿缁涓渚挎峰扳缂杈浠讳哄

CRISPR/Cas9剧ヨ缁锛

CRISPR/Cas绯荤宸ヤ

ｄ锛杩涔瀹崇浣浣

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

CRISPR搴绀烘撅朵腑锛褰㈡琛ㄧず楂搴撮搴锛姝ｆ瑰舰琛ㄧず稿淇瀹澶搴

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

CRISPR/Cas9伐浣绀烘

锛tracrRNA/crRNA浜澶浣瀵Cas9藉惧苟剁瑰DNA锛朵腑查ㄥ涓Cas9斤剧ユ缃缁锛

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

CRISPR/Cas9搴哄查

浠ュ哄查や负渚锛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

CRISPR/Cas9查ゆㄥ哄撅缁捐濯锛

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

CRISPR/Cas9ユ板哄撅缁捐濯锛

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

CRISPR/Cas9ㄥ荤ュ悍浜х娲汇瀹剁茬绛棰搴ㄤ寰浜虹版

CRISPR/Cas9箍锛剧ヨ缁锛

荤ュ悍棰锛浣跨CRISPR归HIV姣璇卞瀹缁姣寤虹妯″绛版Nature绛蹇ラ锛

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

涓杩锛姝ｅ榛芥护浜哄榛锛CRISPR/Cas9旱浣挎璇稿涓纭锛磋ㄦ浠涓杩ヤ╃

绉腑解腑界哄绀句瑰╃ㄤ俊娈靛灞绉瀛绉瀛濞

辩腑借浣哄锛杞浇璇锋敞哄