鍒╃敤CRISPR鎶鏈紝鍚屾椂鏁查櫎涓婂崈涓熀鍥

鍒╃敤鍔熻兘缂哄け鐨勭瓥鐣ヨ繘琛岀殑楂橀氶噺閬椾紶绛涢夛紝鏄爺绌朵汉鍛樺揩閫熷鎵捐皟鎺х壒瀹氳〃鍨嬬殑閲嶈鎴栧叧閿熀鍥犵殑涓昏鏂规硶銆侰RISPR鎶鏈殑鍏磋捣锛屼娇楂橀氶噺绛涢夊熀鍥犳洿鍔犲揩閫熺畝渚匡紝骞惰兘鍑嗙‘鐨勫疄鐜板熀鍥犵粍鏁查櫎绛夌紪杈戝姛鑳斤紝鍥犺屾垚涓轰竴绉嶅己澶х殑閬椾紶绛涢夊伐鍏枫傜浉姣斾箣鍓嶇殑RNAi骞叉壈鐨勬柟娉曪紝搴旂敤CRISPR鎶鏈繘琛岀殑鍩哄洜鏁查櫎鏇村姞绋冲畾锛岀‘淇濆熀鍥犲姛鑳藉畬鍏ㄧ己澶憋紝鏄爺绌朵汉鍛樺揩閫熷彂鐜版柊闈剁偣鏂板熀鍥犵殑閲嶈鏂规硶銆

鍗氶泤杈戝洜鎷ユ湁涓撲笟鐨勯珮閫氶噺閬椾紶绛涢夋妧鏈钩鍙帮紝鍚屾椂鏈変笓涓氱敓鐗╀俊鎭妧鏈汉鍛樿繍鐢ㄦ柊鐨勭畻娉曡璁¢珮鏁堢巼鐨剆gRNA鏂囧簱锛岀粡楠屼赴瀵岀殑瀹為獙鎶鏈汉鍛樹弗鏍兼妸鎺у疄楠岀殑姣忎釜姝ラ锛屽弻鍛ㄦ彁渚涘疄楠岃繘灞曟姤鍛婏紝璁╂偍鏀惧績鐨勬妸瀹為獙浜ょ粰鎴戜滑銆

宸ヤ綔娴佺▼

鍗氶泤杈戝洜浣跨敤娣峰悎鍨嬫枃搴撶瓫閫夌瓥鐣ワ紝鎴戜滑鎷ユ湁涓绯诲垪閽堝浜虹被銆佸皬榧犱互鍙婂叾浠栫墿绉嶇殑sgRNA鏂囧簱锛屼笌姝ゅ悓鏃讹紝鎴戜滑涔熸彁渚涘畾鍒舵湇鍔°傛暣涓瓫閫夌敱浣跨敤鍖呭惈sgRNA鏂囧簱鐨勬參鐥呮瘨渚垫煋缁嗚優寮濮嬶紝鍦ㄥ悎鐞嗘帶鍒朵镜鏌撳鏁帮紙multiplicity of infection锛孧OI锛夌殑鍩虹涓婏紝鎴戜滑鑳藉淇濊瘉姣忎釜琚镜鏌撶殑缁嗚優锛屼粎鏈夊崟涓鐨剆gRNA琚暣鍚堣繘缁嗚優鐨勫熀鍥犵粍銆傚湪渚垫煋瀹屾垚鍚庯紝鎴戜滑浼氫緷鎹墍闇鐮旂┒鐨勯棶棰樺缁嗚優鏂藉姞閫夋嫨鍘嬶紝閫氳繃瀵圭粏鑳炶〃鍨嬬殑瑙傚療瀵圭粏鑳炶繘琛岃仛绫伙紝鍚庣画鍒欏埄鐢ㄦ繁搴︽祴搴忚瀵熶笉鍚岀被鍒粏鑳炵兢鍐卻gRNA涓板害鐨勫彉鍖栵紝鏉ュ垽鏂摢浜涘熀鍥犲弬涓庡湪鎴戜滑鐮旂┒鐨勭敓鐗╅棶棰樹腑銆

screening鈥嬧嬧

CRISPR楂橀氶噺缂栫爜鍩哄洜绛涢夐傜敤浜庯細

  • 瀵绘壘娼滃湪鐨勮嵂鐗╅澏鐐癸紱
  • 绛涢夌浉鍏崇殑鐢熺墿鏍囧織鐗╋紱
  • 绛涢夌粏鑳炴椿鎬с佽嵂鐗╂晱鎰熸ф垨鑰愬彈鍩哄洜锛
  • 鎸囧涓村簥瀹為獙涓殑鍏ョ粍鐥呬汉閫夋嫨锛
  • 鍙戠幇娼滃湪鑱斿悎娌荤枟鐨勯澏鐐规垨淇″彿閫氳矾锛
  • 绛涢夋綔鍦ㄧ殑鍚堟垚鑷存鍩哄洜锛屽紑鍚埗鑽柊鎬濊矾銆

 

妗堜緥灞曠ず锛氬叏鍩哄洜缁勭瓫閫夊彂鐜扮偔鐤藉拰鐧藉枆姣掔礌姣掓ц嚦鍏抽噸瑕佺殑瀹夸富鍩哄洜

 

鐩殑锛閫氳繃鏋勫缓浜虹被鍏ㄥ熀鍥犵粍鏂囧簱锛屽埄鐢ㄦ參鐥呮瘨绯荤粺鏉ユ瀯寤轰汉婧愮粏鑳炴枃搴撶粨鍚堥珮閫氶噺娣卞害娴嬪簭鎶鏈鎵剧偔鐤藉拰鐧藉枆姣掔礌鍦ㄤ汉浣撲腑鐨勫涓诲熀鍥犮

鏂规硶锛

  •  Edi-HTSTM HTSLV60401 鍗氶泤杈戝洜浜虹被鍏ㄥ熀鍥犵粍sgRNA鏂囧簱锛屼汉绫 Whole Human Genome鐥呮瘨鏂囧簱Edi-HTSTM WHG鏄痵gRNA杞戒綋鏂囧簱锛屾诲叡225484鏉gRNA锛屽叾涓瘡涓紪杈戝熀鍥犲潎鏈 6-10 涓澏鐐 sgRNA锛 鍩哄洜鏁拌揪鍒颁簡 18823 涓熀鍥犮侲di-HTSTM WHG 鏂囧簱鍙互鍦ㄤ汉绫荤粏鑳炲唴鏁查櫎鎵鏈夌殑 Whole Human Genome 鐩稿叧缂栫爜鍩哄洜锛
  • 鍒╃敤Edi-HTSTM WHG鏂囧簱鏋勫缓HeLa缁嗚優鏂囧簱锛
  • 灏哖A/LFnDTA鍜宒iphtheria toxin (DT)鍔犲叆鍒扮粏鑳炴枃搴撲腑杩涜绛涢夛紱
  • NGS妫娴嬬瓫閫夊悗鐨勭粏鑳烇紝杩涜鏁版嵁鍒嗘瀽锛屾壘鍒版綔鍦ㄩ澏鐐广

 

2 1

鍥2 鐐柦鍜岀櫧鍠夋瘨绱犵瓫閫夊悗鐨勫叧閿涓诲熀鍥

 CRISPR楂橀氶噺LncRNA绛涢


Reference

1. Zhou Y, Zhu S, Cai C, Yuan P, Li C, Huang Y and Wei W*. (2014) High-throughput Screening of a CRISPR/Cas Library for Functional Genomics in Human Cells. Nature 509(7501): 487-91.

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