7.17<\/p>\n\n\n\n
\u524d\u4e00\u5929\u73a9\u624b\u673a\u901a\u5bb5\u4e86\uff0c\u4e00\u4e0b\u90fd\u6ca1\u7761\uff0c\u65e9\u4e0a\u4e5d\u70b9\u591a\u4e00\u70b9\u5230\u4e86\u5b9e\u9a8c\u5ba4\uff0c\u68c0\u67e5\u628atRNA ligation complex\u7684\u8d28\u7c92GFP\u6362\u6210tagBFP\u7684\u6d4b\u5e8f\u7ed3\u679c\uff0c\u53d1\u73b0\u90fd\u4e0d\u5bf9\uff0c\u60f3\u8d77\u6765\u662f\u56e0\u4e3a\u81ea\u5df1\u5c11\u505a\u4e86\u4e00\u6b65\u9176\u5207\u3002\u7136\u540e\u53c8\u91cd\u65b0\u9176\u5207\u3001\u8fde\u63a5\u3001\u8f6c\u5316\u3002\u665a\u4e0a\u5341\u4e8c\u70b9\u6765\u6311\u83cc\u83cc\u843dPCR\u3002<\/p>\n\n\n\n
\u4e2d\u5348\u6ca1\u5403\u996d\uff0c\u76f4\u63a5\u5728\u5b9e\u9a8c\u5ba4\u6253\u4e86\u4e00\u628a\u7ea2\u8b66\uff0c\u7136\u540e\u8db4\u684c\u5b50\u4e0a\u7761\u4e86\u56db\u4e2a\u5c0f\u65f6\u3002\u9192\u7684\u65f6\u5019\u53d1\u73b0\u809a\u5b50\u5de8\u997f\u65e0\u6bd4\uff0c\u7136\u540e\u53bb\u4e07\u4eba\u5403\u4e86\u4e2a\u996d\uff0c\u5403\u5b8c\u5f00\u59cb\u80c3\u75bc\uff0c\u75bc\u5230\u76f4\u4e0d\u8d77\u8170\u3002\u4e8e\u662f\u653e\u5f03\u4e86\u505a\u5b8cSP6\u542f\u52a8\u5b50\u63a7\u5236\u7684VPR-dCas9\u548cKRAB-dCas9\u7684\u4f53\u5916\u8f6c\u5f55\u7684\u6253\u7b97\u3002\u6b63\u5de7\u89c2\u5bdfSalI\u5bf9\u8d28\u7c92\u7ebf\u6027\u5316\u7684\u7ed3\u679c\uff0c\u53d1\u73b0\u661f\u53f7\u6d3b\u6027\u6781\u5176\u663e\u8457\uff0c\u8d28\u7c92\u90fd\u6ca1\u4e3b\u5e26\u4e86\u3002\u5c31\u6254\u6389\u4e86\u3002<\/p>\n\n\n\n
\u56de\u5bbf\u820d\u8eba\u5e8a\u4e0a\u4e00\u4f1a\u513f\uff0c\u809a\u5b50\u5c31\u4e0d\u75db\u4e86\u3002\u6574\u7406\u4e86\u4e00\u4e0b\u9f99\u821f\u961f\u6691\u671f\u6253\u5361\u7b2c\u4e00\u5468\u5927\u5bb6\u7684\u6253\u5361\u60c5\u51b5\u3002\u7136\u540e\u5728\u5e8a\u4e0a\u770b\u4e86\u4e24\u96c6\u4e09\u4f53\u7535\u89c6\u5267\u3002\u7136\u540e\u70b9\u4e86\u4e00\u70b9\u6c34\u679c\u5916\u5356\u5403\u6389\u4e86\u3002<\/p>\n\n\n\n
\u7136\u540e\u53c8\u56de\u5b9e\u9a8c\u5ba4\uff0c\u770b\u770b\u4e3a\u5565\u8d28\u7c92\u6ca1\u6709\u4e3b\u5e26\uff0c\u8fd9\u6b21\u52a0\u4e86\u73af\u5f62\u5bf9\u7167\u3002\u53d1\u73b0\u6761\u5e26\u5f88\u5355\u4e00\uff0c\u6362\u4e86KpnI\u5355\u9176\u5207\uff0c\u8fd8\u6709\u6709\u661f\u53f7\u6d3b\u6027\u3002\u53ef\u80fd\u662f\u56e0\u4e3a\u8fd9\u6b21\u63d0\u8d28\u7c92\u7684\u65f6\u5019\u7528\u4e86100\u03bcl\u6c34\u6d17\u8131\uff0c\u4f53\u7cfb\u91cc\u76d0\u79bb\u5b50\u6bd4\u8f83\u591a\uff1f\u73b0\u5728\u4e5f\u6ca1\u5b9a\u8bba\u3002<\/p>\n\n\n\n
\u7136\u540e\u5c31\u50cf\u5148\u91cd\u65b0\u63d0\u4e2a\u8d28\u7c92\uff0c\u83cc\u6db2\u4e0d\u5728\u4e86\u3002\u505a\u4e86\u4e2a\u8f6c\u5316\u3002<\/p>\n\n\n\n
7.18<\/p>\n\n\n\n
\u524d\u4e00\u5929\u7684\u4e1c\u897f\u505a\u5b8c\u5df2\u7ecf\u662f7\u670818\u65e5\u65e9\u4e0a5\u70b9\u4e86\u3002\u5199\u4e86\u4e2abiweek work report\uff0c\u7136\u540e\u56de\u5bdd\u5ba4\u7761\u89c9\u3002<\/p>\n\n\n\n
7.19<\/p>\n\n\n\n
\u665a\u4e0a\u7236\u4eb2\u6709\u7535\u8bdd\u6253\u8fc7\u6765\u3002\u4e00\u5982\u65e2\u5f80\u7684\u4e0d\u77e5\u9053\u8bf4\u5565\u3002<\/p>\n\n\n\n
7.20<\/p>\n\n\n\n
\u665a\u4e0a\u5728\u4e1c\u64cd\u8dd1\u5eca\u6709\u4eba\u4ed6\u4eec\u7ed9\u6211\u4e70\u4e86\u86cb\u7cd5\uff0c\u731c\u6d4b\u662f\u60e0\u60e0\u544a\u8bc9\u4e86\u5e2d\u6cfd\uff0c\u7136\u540e\u4e2d\u9014\u6211\u8fd8\u770b\u5230\u5e2d\u6cfd\u51fa\u53bb\u4e86\u4e00\u8d9f\u3002\u53ef\u80fd\u662f\u53bb\u62ff\u86cb\u7cd5\u3002<\/p>\n\n\n\n
\u665a\u4e0a\u9664\u51b0\u592a\u723d\u4e86\uff0c\u867d\u7136\u6211\u77e5\u9053\u7b49\u5b83\u5316\u4e00\u665a\u4e0a\u5c31\u597d\u4e86\uff0c\u4f46\u8fd8\u662f\u5fcd\u4e0d\u4f4f\u7528\u6273\u624b\u3001\u87ba\u4e1d\u5200\u548c\u94c1\u68d2\u6572\u4e86\u4e24\u4e2a\u5c0f\u65f6\u3002\u4e09\u70b9\u591a\u624d\u5f04\u5b8c\u3002<\/p>\n\n\n\n
7.21<\/p>\n\n\n\n
\u65e9\u4e0a\u5341\u70b9\u5230\u7684lab\uff0c\u5403\u4e86\u4e00\u6876\u6ce1\u9762\u52a0\u706b\u817f\u80a0\uff0c\u7136\u540e\u63d0\u4e86\u4e2aL1-KO\u7684RNA\uff0c\u672c\u6765\u8bf4\u4e0b\u5348\u4e24\u70b9\u534a\u505aqPCR\uff0c\u5e08\u59d0\u89c9\u5f97\u8d76\u4e0d\u4e0a\uff0c\u4e8e\u662f\u6211\u653e\u4e8675%\u4e59\u9187\u540e\u5c31\u53bb\u5403\u996d\u4e86\uff0c\u5403\u5b8cZT\u56delab\u505a\u5b9e\u9a8c\uff0c\u6211\u56de\u5bdd\u5ba4\u7761\u89c9\u4e86\uff0c\u4e00\u4e0b\u7761\u4e86\u56db\u4e2a\u591a\u5c0f\u65f6\u3002\u5230\u5b9e\u9a8c\u5ba4\u5f00\u7ec4\u4f1a\u3002<\/p>\n\n\n\n
7.22<\/p>\n\n\n\n
\u6211\u597d\u50cf\u4e24\u5468\u540e\u5c31\u8981\u8bb2\u5de5\u4f5c\u8fdb\u5c55\u4e86\uff0c\u6ca1\u7684\u8bf4\u5440\u3002<\/p>\n\n\n\n
\u65e9\u4e0a\u9192\u7684\u5f88\u65e9\uff0c\u7136\u540e\u770b\u5916\u9762\u4e0b\u96e8\u7279\u522b\u5927\uff0c\u6211\u53c8\u6ca1\u4f1e\uff0c\u7ea0\u7ed3\u4e86\u597d\u4e45\uff0c\u9876\u7740\u4e00\u4e2a\u7eb8\u7bb1\u5b50\u51fa\u53bb\u4e86\uff0c\u8fd8\u597d\u7a7f\u4e86\u62d6\u978b\uff0c\u8e5a\u6c34\u597d\u51e0\u6b21\u3002\u65e9\u4e0a\u505a\u5b8c\u53cd\u8f6c\uff0c\u4e0b\u5348\u505a\u4e86qPCR\uff0cLY\u5e08\u59d0\u6559\u6211\u3002\u7136\u540e\u67e5\u4e86\u67e5FANA ASO\u548cMorpholino ASO\u7684\u533a\u522b\u3002\u665a\u4e0a\u53bb\u897f\u4f53\u8bad\u7ec3\uff0c\u7ec3\u5b8c\u5403\u4e86\u4e2a\u996d\u7136\u540e\u56delab\u6784\u5efaPB-EF1a-tagBFP-T2A-HA-RTCB\/DDX1\/CGI-99\/FAM98B\/ASW.<\/p>\n\n\n\n
7.23<\/p>\n\n\n\n
\u6784\u5b8c\u5c31\u65e9\u4e0a\u516d\u70b9\u4e86\uff0c\u56de\u53bb\u7761\u5230\u4e0b\u5348\u3002\u53bb\u5b9e\u9a8c\u5ba4\u5904\u7406\u4e86\u4e00\u4e0bqPCR\u7684\u6570\u636e\uff0c\u7136\u540e\u665a\u4e0a\u6311\u83cc\u9a8c\u8bc1\u3002\u56de\u5bbf\u820d\u662f\u4e00\u70b9\u3002<\/p>\n\n\n\n
7.24<\/p>\n\n\n\n
\u665a\u4e0a\u53ea\u7761\u4e86\u4e24\u4e2a\u534a\u5c0f\u65f6\u5c31\u81ea\u7136\u9192\u4e86\uff0c\u7136\u540e\u4e0a\u4e86\u4e2a\u5395\u6240\u5c31\u7761\u4e0d\u7740\u4e86\u3002\u770b\u4e86\u4e00\u7ae0\u897f\u65b9\u54f2\u5b66\u53f2\uff08\u5965\u53e4\u65af\u4e01\uff09\u3002\u7136\u540e\u53bb\u5b9e\u9a8c\u5ba4\u6253\u4e86\u4e24\u628aCF\uff0c\u7136\u540e\u53bb\u5403\u4e86\u4e2a\u65e9\u996d\u3002<\/p>\n\n\n\n
7.25~7.27\u5fd8\u4e86\u5e72\u5565\u4e86\uff0c\u53ea\u8bb0\u5f97\u6784\u5efa\u597d\u4e86\u8def\u579a\u5e08\u59d0\u591f\u4e86\u51e0\u4e2a\u661f\u671f\u6ca1\u5f04\u51fa\u6765\u7684PX320-GFP. PB-EF1a-tagBFP-T2A-HA-RTCB\/DDX1\/CGI-99\/FAM98B\/ASW\u6d4b\u5e8f\u4e5f\u5bf9\u4e86\uff0c\u505a\u4e86\u9187\u6c89\u3002<\/p>\n\n\n\n
7.28<\/p>\n\n\n\n
\u665a\u4e0a\u8981\u5f00\u7ec4\u4f1a\u624d\u5230\u5b9e\u9a8c\u5ba4\uff0c\u4e9a\u5f3a\u5e08\u5144\u8bb2\u4e86\u4e00\u4e0bSAFB\u7684\u8bba\u6587\u7684\u4fee\u8865\uff0c\u7136\u540e\u5c0f\u6db5\u5e08\u59d0\u8bb2\u4e86\u4e00\u7bc7\u6211\u6ca1\u770b\u4e5f\u542c\u5979\u4e0d\u61c2\u7684enhancer\u76f8\u5173\u7684\u6587\u732e\u3002\u53f2\u4e0a\u6700\u5feb\u7ec4\u4f1a\u3002\u7ec4\u4f1a\u7ed3\u675f\u540e\uff0c\u5f80PX320-GFP\u91cc\u52a0\u4e8610\u4e2asgRNA\uff0c\u5149\u8001\u5e08\u4e5f\u505a\u5b9e\u9a8c\u5230\u633a\u665a\u3002\u505a\u5b8c\u6211\u6253\u4e86\u4e24\u628aCF\u3002\u4e0b\u5468\u7ec4\u4f1a\u5c31\u662f\u6211\u4e86\uff0c\u53ef\u662f\u4e0d\u77e5\u9053\u8be5\u8bb2\u4ec0\u4e48\u3002<\/p>\n\n\n\n
7.29<\/p>\n\n\n\n
\u65e9\u4e0a\u4e03\u70b9\u5c31\u5230\u5b9e\u9a8c\u5ba4\u4e86\uff0c\u867d\u7136\u524d\u4e00\u5929\u665a\u4e0a\u56de\u53bb\u5df2\u7ecf\u51cc\u6668\u4e00\u70b9\u4e86\u3002\u7761\u4e86\u4e00\u4e2a\u534a\u5c0f\u65f6\u5c31\u9192\u4e86\uff0c\u5fd8\u8bb0\u662f\u56e0\u4e3a\u70ed\u8fd8\u662f\u56e0\u4e3a\u868a\u5b50\u4e86\u3002\u7136\u540e\u5c31\u7761\u4e0d\u7740\u4e86\uff0c\u5728\u5e8a\u4e0a\u770b\u4e86\u4e00\u7ae0\u897f\u65b9\u54f2\u5b66\u53f2\uff0c\u7136\u540e\u7b49\u5230\u516d\u70b9\u591a\u6709\u70ed\u6c34\u4e86\u6d17\u4e86\u4e2a\u6fa1\uff0c\u53bb\u4e01\u9999\u56ed\u5403\u4e86\u4e2a\u65e9\u996d\u3002\u5230\u5b9e\u9a8c\u5ba4\u597d\u50cf\u6ca1\u5e72\u5565\u4e8b\uff0c\u53ef\u80fd\u9274\u5b9a\u4e86\u4e00\u4e0b\u8f6c\u5316\u5b50\uff0c\u7528sgRNA\u7684\u5f15\u7269\u9274\u5b9a\u7684\uff0c\u8fd8\u633a\u7ba1\u7528\u3002<\/p>\n\n\n\n
\u5403\u4e86\u5348\u996d\u56de\u5bbf\u820d\u7761\u4e86\u56db\u4e2a\u5c0f\u65f6\uff0c\u9192\u540e19\u70b9\u8ddf\u8c2d\u4eba\u8c6a\u4e24\u4e2a\u4eba\u53bb\u897f\u4f53\u7ec3\u4e86\u80a9\u80cc\u3002\u6211\u53c8\u8ddf\u4ed6\u63d0\u4e86\u65b0\u751f\u5165\u5b66\u7684\u65f6\u5019\u53bb\u7a7f\u7740\u961f\u670d\u642c\u884c\u674e\u7684\u4e8b\u60c5\uff0c\u4ed6\u4e0d\u662f\u5f88\u613f\u610f\u3002<\/p>\n\n\n\n
\u51cc\u6668\u4e00\u70b9\u7684\u65f6\u5019\u6447\u4e86PX320-GFP-sg1~10\u83cc\u3002\u7136\u540e\u8fd9\u4e00\u5929\u8fd8\u7cfb\u7edf\u7684\u6574\u7406\u4e86\u4e00\u4e0b\u6db2\u6c2e\u7f50\u548c\u8d28\u7c92\uff0c\u5efa\u7acb\u4e86\u7d22\u5f15\u8868\uff0c\u76d2\u5b50\u4e0a\u6807\u597d\u4e86\u4f4d\u7f6e\u7684\u5e8f\u53f7\u3002<\/p>\n\n\n\n
7.30<\/p>\n\n\n\n
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