{"id":26830,"date":"2017-02-24T00:13:01","date_gmt":"2017-02-23T16:13:01","guid":{"rendered":"https:\/\/case.ntu.edu.tw\/blog\/?p=26830"},"modified":"2018-07-29T00:37:15","modified_gmt":"2018-07-28T16:37:15","slug":"%e7%99%bb%e4%b8%8a-crispr-%e6%b5%aa%e6%bd%ae","status":"publish","type":"post","link":"https:\/\/case.ntu.edu.tw\/blog\/?p=26830","title":{"rendered":"\u767b\u4e0a CRISPR \u6d6a\u6f6e"},"content":{"rendered":"
\"Caroline
Caroline Davis2010 | Flickr<\/figcaption><\/figure>\n

\u64b0\u6587\uff5c<\/strong>\u99ac\u5343\u60e0<\/p>\n

\u5c0d\u5206\u5b50\u751f\u7269\u5b78\u5bb6\u800c\u8a00\uff0cCRISPR \u662f\u4e00\u975e\u5e38\u5f37\u5927\u7684\u5de5\u5177\uff0c\u53ef\u4ee5\u7528\u4f86\u66f4\u77ad\u89e3\u57fa\u56e0\u9ad4\u5982\u4f55\u904b\u4f5c\uff0c\u4ee5\u53ca\u63a2\u8a0e\u66f4\u6df1\u5165\u7684\u554f\u984c\u30022016\u5e74\u300c\u81ea\u7136\u300d\u671f\u520a\uff08Nature \uff09\u6574\u7406\u51fa CRISPR-Cas9 \u767c\u5c55\u7684\u4e94\u500b\u65b9\u5411\uff0c\u53ca\u5176\u5982\u4f55\u6539\u8b8a\u79d1\u5b78\u5bb6\u5011\u7814\u7a76\u7684\u65b9\u6cd5\u3002<\/p>\n

\uff08\u4e00\uff09\u58de\u6389\u7684\u526a\u5200\uff08BROKEN SCISSORS)<\/strong><\/span><\/p>\n

CRISPR-Cas9 \u7cfb\u7d71\u4e3b\u8981\u7531\u5169\u500b\u90e8\u5206\u7d44\u6210\uff0c\u4e00\u500b\u53ef\u4ee5\u8fa8\u8b58\u7279\u5b9a\u57fa\u56e0\u5e8f\u5217\u7684\u5f15\u5c0eRNA \u5206\u5b50\uff0c\u548c\u4e00\u500b\u5177\u6709\u5206\u5b50\u526a\u5200\u7279\u6027\u7684 Cas9 \u9175\u7d20\uff0c\u5f15\u5c0eRNA \u5206\u5b50\u8b93 Cas9 \u9175\u7d20\u8207\u7279\u5b9a\u57fa\u56e0\u5e8f\u5217\u7d50\u5408\uff0c\u9032\u800c\u526a\u65b7DNA\u3002\u6b64\u7279\u6027\u63d0\u4f9b\u60f3\u8981\u85c9\u7531\u7834\u58de\u7279\u5b9a\u57fa\u56e0\u4f86\u7814\u7a76\u5176\u529f\u80fd\u7684\u79d1\u5b78\u5bb6\u4e00\u500b\u5a01\u529b\u5f37\u5927\u7684\u5de5\u5177\u3002\u540c\u6642\u914d\u5408\u63d0\u4f9b\u7279\u5b9a\u7684\u4fee\u5fa9\u6a21\u7248\uff0c\u4f7f\u57fa\u56e0\u4ee5\u4e0d\u540c\u65b9\u5f0f\u4fee\u5fa9\uff0c\u53ef\u4ee5\u8b93\u7814\u7a76\u8005\u5728\u4efb\u4f55\u57fa\u56e0\u9ad4\u7279\u5b9a\u4f4d\u7f6e\u653e\u5165\u4efb\u4f55\u4e00\u6bb5\u65b0\u5e8f\u5217\u3002<\/p>\n

2012\u5e74\uff0c\u7576\u5927\u5bb6\u6b63\u81f4\u529b\u65bc\u5229\u7528\u6b64\u5de5\u5177\u526a\u65b7\u4eba\u985e\u57fa\u56e0\u6642\uff0c\u52a0\u5dde\u5927\u5b78\u820a\u91d1\u5c71\u5206\u6821\u7cfb\u7d71\u751f\u7269\u5b78\u5bb6Jonathan Weissman \u6240\u5e36\u9818\u7684\u5718\u968a\u537b\u6709\u4e0d\u4e00\u6a23\u7684\u505a\u6cd5: \u7834\u58de\u5206\u5b50\u526a\u5200\u3002Stanley Qi \u5c07 Cas9 \u9175\u7d20\u52a0\u4ee5\u7a81\u8b8a\uff08mutate\uff09\uff08\u8a3b1\uff09\uff0c\u4f7f\u5176\u96d6\u7136\u53ef\u4ee5\u7d50\u5408\u81f3\u5f15\u5c0e RNA \u6240\u8fa8\u8b58\u7684\u57fa\u56e0\u5e8f\u5217\uff0c\u537b\u5931\u53bb\u5c07\u5176\u5207\u65b7\u7684\u529f\u80fd\uff0c\u540c\u6642\u8a98\u8b8a\u7684 Cas9\u00a0 \u9175\u7d20\u963b\u64cb\u4e86\u5176\u4ed6\u8f49\u9304\u6240\u9700\u7684\u86cb\u767d\u8cea\u8207\u6b64\u7279\u5b9a\u57fa\u56e0\u7d50\u5408\uff0c\u5f62\u540c\u5728\u4e0d\u5f71\u97ff\u8207\u6539\u8b8aDNA \u5e8f\u5217\u7684\u72c0\u6cc1\u4e0b\u95dc\u6389\u6b64\u7279\u5b9a\u57fa\u56e0\u7684\u57fa\u56e0\u8868\u73fe\u3002\u79d1\u5b78\u5bb6\u4e0d\u6eff\u8db3\u6b62\u6b65\u65bc\u6b64\uff0c\u4ed6\u5011\u66f4\u9032\u4e00\u6b65\u518d\u5c07\u53ef\u4ee5\u6d3b\u5316\u57fa\u56e0\u8868\u73fe\u7684\u86cb\u767d\u8cea\u63a5\u5230\u9019\u88ab\u7834\u58de\u7684\u526a\u5200 \uff08\u8a98\u8b8aCas9 \u9175\u7d20\uff09\u4e0a\uff0c\u8b93\u7279\u5b9a\u57fa\u56e0\u8868\u73fe\u5f97\u4ee5\u88ab\u6d3b\u5316\uff0c\u5efa\u7acb\u4e86\u4e00\u5957\u5728\u4e0d\u5f71\u97ff\u539fDNA \u5e8f\u5217\u72c0\u6cc1\u4e0b\uff0c\u53ef\u4ee5\u81ea\u7531\u6253\u958b\u6216\u95dc\u9589\u57fa\u56e0\u8868\u73fe\u7684\u7cfb\u7d71\u3002\u6578\u500b\u5be6\u9a57\u5ba4\u5718\u968a\u4e5f\u9678\u7e8c\u767c\u8868\u4e86\u66f4\u591a\u6210\u679c\uff0c\u751a\u81f3\u53ef\u5728\u540c\u4e00\u5be6\u9a57\u4e2d\u8abf\u63a7\u4e09\u500b\u6216\u66f4\u591a\u7684\u57fa\u56e0\uff0c\u61c9\u7528\u65bc\u8907\u96dc\u751f\u7269\u6a5f\u5236\u7684\u7814\u7a76\u3002<\/p>\n

(\u4e8c\uff09CRISPR \u975e\u907a\u50b3\u6027\u7684\u8abf\u63a7 (CRISPR EPIGENETICS)<\/strong><\/span><\/p>\n

\u5f9e\u4e8b\u57fa\u56e0\u7642\u6cd5\u7814\u7a76\u7684\u8377\u862d\u907a\u50b3\u5b78\u5bb6 Marianne Rot\uff0c\u4e00\u958b\u59cb\u81f4\u529b\u65bc\u5c0b\u627e\u9020\u6210\u75be\u75c5\u7684\u7a81\u8b8a\u57fa\u56e0\uff0c\u4f46\u662f\u5f8c\u4f86\u767c\u73fe\u66f4\u591a\u75be\u75c5\u662f\u4f86\u81ea\u57fa\u56e0\u8868\u73fe\u53d7\u5230\u5e72\u64fe\uff0c\u800c\u975e\u6e90\u65bc\u55ae\u4e00\u7684\u57fa\u56e0\u7a81\u8b8a\u3002\u5979\u8a8d\u70ba\u6700\u597d\u63a7\u5236\u57fa\u56e0\u6d3b\u6027\u7684\u65b9\u6cd5\uff0c\u5c31\u662f\u8abf\u63a7\u57fa\u56e0\u8868\u73fe\u3002\u7d44\u86cb\u767d\uff08histones) \u662f\u7e8f\u7e5e\u5305\u88f9DNA \u7684\u86cb\u767d\u8cea\uff0c\u7d44\u86cb\u767d\u5305\u88f9 DNA \u7684\u7a0b\u5ea6\u6703\u5f71\u97ff\u5230\u5176\u4ed6\u5230\u86cb\u767d\u8cea\u9760\u8fd1DNA\u7684\u60c5\u5f62\uff0c\u4e14\u7d44\u86cb\u767d\u8207DNA\u7684\u7d50\u5408\u7a0b\u5ea6\u6703\u96a8\u8457\u500b\u9ad4\u767c\u80b2\u548c\u74b0\u5883\u800c\u6539\u8b8a\u3002\u79d1\u5b78\u5bb6\u767c\u73fe\uff1a\u5f9e\u5927\u8166\u6d3b\u52d5\u5230\u816b\u7624\u751f\u9577\uff0c\u90fd\u548c\u57fa\u56e0\u5e8f\u5217\u4ee5\u5916\u7684\u8abf\u63a7\u6709\u95dc\uff0c\u4f46\u662f\u7814\u7a76\u7684\u74f6\u9838\u5728\u65bc\u7121\u6cd5\u6539\u8b8a\u7279\u5b9a\u57fa\u56e0\u4f4d\u7f6e\u7684\u6a19\u8a18\u3002CRISPR-Cas9 \u6280\u8853\u7684\u51fa\u73fe\uff0c\u8b93\u9019\u500b\u7814\u7a76\u74f6\u9838\u770b\u5230\u89e3\u5957\u7684\u66d9\u5149\u30022015\u5e74Charles Gersbach \u6210\u529f\u5730\u5c07\u4e00\u500b\u53ef\u4ee5\u5c07\u4e59\u91af\u57fa\u63a5\u5230\u7d44\u86cb\u767d\u4e0a\u7684\u9175\u7d20\u63a5\u5728\u5931\u53bb\u526a\u65b7 DNA \u529f\u80fd\u7684 Cas9 \u9175\u7d20\u4e0a\uff0c\u518d\u85c9\u7531\u5f15\u5c0e RNA \u5c07\u6b64\u9175\u7d20\u5e36\u81f3\u76ee\u6a19\u57fa\u56e0\uff0c\u9032\u800c\u5f71\u97ff\u8a72\u57fa\u56e0\u7684\u8868\u73fe\u3002<\/p>\n

\uff08\u4e09\uff09CRISPR \u7834\u89e3\u5bc6\u78bc (CRISPR CODE CRACKING)<\/strong><\/span><\/p>\n

\u8d85\u904e98%\u7684\u4eba\u985eDNA\u548c\u86cb\u767d\u8cea\u7684\u5408\u6210\u7121\u95dc\uff0c\u79d1\u5b78\u5bb6\u5011\u8a8d\u70ba\u9019\u5927\u591a\u6578\u7684DNA \u61c9\u8a72\u4e5f\u6709\u91cd\u8981\u529f\u80fd\uff0c\u53ea\u662f\u6211\u5011\u9084\u4e0d\u77e5\u9053\u3002CRISPR-Cas9 \u6280\u8853\u63d0\u4f9b\u4e86\u4e00\u500b\u65b9\u6cd5\u4f86\u63a2\u8a0e\u6b64\u4e00\u554f\u984c\u3002\u5229\u7528\u6b64\u6280\u8853\uff0c\u79d1\u5b78\u5bb6\u5011\u53ef\u4ee5\u91dd\u5c0d\u7279\u5b9a\u5e8f\u5217\uff0c\u4f8b\u5982\u505a\u70ba\u5897\u5f37\u5b50\uff08enhancer) \u6216\u8f49\u9304\u51fa\u975e\u8f49\u8b6f\u578b RNA \u7684 DNA \u5e8f\u5217\u53bb\u505a\u7814\u7a76\u3002Farnham \u548c\u5979\u7684\u5718\u968a\u767c\u73fe\uff0c\u539f\u672c\u4ee5\u70ba\u548c\u524d\u5217\u817a\u764c\u548c\u5927\u8178\u764c\u6709\u95dc\u7684\u7a81\u8b8a\u52a0\u5f37\u5b50\uff0c\u7d93\u7531CRISPR-Cas9 \u522a\u9664\u5f8c\uff0c\u4e26\u4e0d\u5f71\u97ff\u65c1\u908a\u57fa\u56e0\u7684\u8868\u73fe\uff0c\u986f\u793a\u6211\u5011\u539f\u672c\u7528\u4f86\u5206\u6790\u52a0\u5f37\u5b50\u5f37\u5ea6\u7684\u65b9\u6cd5\uff0c\u4e26\u4e0d\u80fd\u53cd\u6620\u51fa\u771f\u5be6\u72c0\u6cc1\u3002\u7531\u907a\u50b3\u5b78\u5bb6David Gifford \u548cRichard Sherwood \u9818\u5c0e\u7684\u7814\u7a76\u5718\u968a\u5728\u56db\u842c\u9e7c\u57fa\u5c0d\u9577\u7684 DNA \u5e8f\u5217\u505a\u4e00\u7cfb\u5217\u7684\u7a81\u8b8a\uff0c\u5617\u8a66\u5b9a\u51fa\u5f71\u97ff\u57fa\u56e0\u8868\u73fe\u7684 DNA \u5e8f\u5217\u5716\u8b5c\u3002\u4e0d\u904e CRISPR-Cas9 \u6280\u8853\u4e5f\u4e0d\u662f\u842c\u80fd\u7684\uff0c\u5728\u6b64\u65b9\u9762\u7684\u61c9\u7528\u6709\u5176\u4fb7\u9650\u6027\uff0c\u7562\u7adf Cas9 \u9175\u7d20\u53ea\u80fd\u7531\u5f15\u5c0e RNA \u5e36\u81f3\u7279\u5b9a\u5340\u57df\uff0c\u4e14\u5728\u9760\u5207\u5272\u4f4d\u7f6e\u8655 DNA\u5e8f\u5217\u5fc5\u9808\u6709\u4e00\u5b9a\u9e7c\u57fa\u5c0d\u5e8f\u5217\u5171\u901a\u6027\uff0c\u7576\u9032\u884c\u975e\u8f49\u8b6f\u578b DNA\u7684\u7814\u7a76\u6642\uff0c\u4e0d\u80fd\u5728DNA\u4efb\u4f55\u4f4d\u7f6e\u4efb\u610f\u5207\u5272\uff0c\u56e0\u70ba\u53ef\u80fd\u6703\u7522\u751f\u5404\u7a2e\u975e\u9810\u671f\u7684\u5f71\u97ff\u3002\u6700\u8fd1\u5f35\u92d2\u6240\u5e36\u9818\u7684\u5718\u968a\u627e\u5230\u53e6\u4e00\u7fa4 Cpf1 \u9175\u7d20\u7fa4\uff0c\u5176\u4f5c\u7528\u548c Cas9\u00a0 \u9175\u7d20\u985e\u4f3c\uff0c\u4f46\u662f\u53ef\u8fa8\u8b58\u4e0d\u4e00\u6a23\u7684DNA\u5e8f\u5217\uff0c\u6216\u8a31\u672a\u4f86\u53ef\u4ee5\u767c\u73fe\u5404\u7a2e\u4e0d\u540c\u9175\u7d20\uff0c\u7528\u4f86\u5207\u5272\u6240\u6709\u60f3\u8981\u7814\u7a76\u7684\u57fa\u56e0\u9ad4\u4f4d\u7f6e\u3002<\/p>\n

\uff08\u56db\uff09CRISPR \u4ee5\u5149\u4f86\u63a7\u5236\u57fa\u56e0\u8868\u73fe (CRISPR SEES THE LIGHT)<\/strong><\/span><\/p>\n

Gersbach\u7684\u5be6\u9a57\u5ba4\u4f7f\u7528\u57fa\u56e0\u7de8\u8f2f\u6280\u8853\u4f86\u63a2\u8a0e\u7d30\u80de\u547d\u904b\u4ee5\u53ca\u5176\u8abf\u63a7\uff0c\u4ed6\u5011\u5e0c\u671b\u80fd\u5728\u57f9\u990a\u76bf\u88cf\u57f9\u990a\u7d44\u7e54\uff0c\u7528\u4f86\u7be9\u6aa2\u85e5\u7269\uff0c\u4e26\u6e2c\u8a66\u7d30\u80de\u6cbb\u7642\u6210\u6548\u3002\u4f46\u662f CRISPR-Cas9 \u7684\u4f5c\u7528\u7d50\u679c\u662f\u6c38\u4e45\u6027\u7684\uff0c\u8207\u4ed6\u5011\u9700\u8981\u80fd\u5920\u66ab\u6642\u6253\u958b\u6216\u95dc\u6389\u57fa\u56e0\u8868\u73fe\u7684\u671f\u5f85\u4e0d\u5408\u3002\u65bc\u662f\u4ed6\u5011\u52a0\u4e86\u4e00\u500b\u80fd\u88ab\u85cd\u5149\u6d3b\u5316\u7684\u86cb\u767d\u8cea\u5230\u5931\u53bb\u5207\u5272\u80fd\u529b\u7684 Cas9 \u9175\u7d20\u4e0a\uff1a\u7576\u4ed6\u5011\u7528\u85cd\u5149\u7167\u5c04\u7d30\u80de\uff0c\u7279\u5b9a\u57fa\u56e0\u958b\u59cb\u8868\u73fe\uff0c\u95dc\u6389\u85cd\u5149\u7279\u5b9a\u57fa\u56e0\u5247\u505c\u6b62\u8868\u73fe\u3002\u5176\u4ed6\u5718\u968a\u4e5f\u6709\u767c\u5c55\u51fa\u985e\u4f3c\u7684\u6280\u8853\uff0c\u6539\u4ee5\u5316\u5b78\u7269\u8cea\u7576\u4f5c\u57fa\u56e0\u8868\u73fe\u7684\u958b\u95dc\u3002<\/p>\n

\uff08\u4e94\uff09\u4ee5CRISPR\u4f86\u88fd\u9020\u5404\u7a2e\u751f\u7269\u6a21\u578b\uff08MODEL CRISPR)<\/strong><\/span><\/p>\n

\u4ee5\u539f\u6709\u7684\u6280\u8853\u8981\u88fd\u9020\u51fa\u57fa\u56e0\u8f49\u6b96\u52d5\u7269\u4f86\u4f5c\u70ba\u7814\u7a76\u6a21\u578b\u662f\u975e\u5e38\u8017\u6642\u8017\u8cbb\u7684\u5de5\u7a0b\uff0cCRISPR-Cas9 \u5c07\u9019\u4ee5\u5e74\u70ba\u55ae\u4f4d\u7684\u904e\u7a0b\u5927\u5e45\u7e2e\u77ed\u70ba\u4e00\u500b\u6708\u5167\u53ef\u5b8c\u6210\u7684\u5be6\u9a57\u3002\u6240\u4ee5\u8a31\u591a\u4e4b\u524d\u7121\u6cd5\u505a\u5230\u7684\u7814\u7a76\uff0c\u5982\u4eca\u90fd\u6709\u53ef\u80fd\u9054\u6210\uff0c\u4f8b\u5982\u5f9e\u764c\u75c7\u5230\u795e\u7d93\u9000\u5316\u7814\u7a76\uff0c\u90fd\u5df2\u63a1\u7528\u6b64\u4e00\u65b0\u6280\u8853\u3002<\/p>\n

\u4e00\u822c\u800c\u8a00\uff0c\u7814\u7a76\u55ae\u4f4d\u6703\u5c07\u4ed6\u5011\u65b0\u767c\u5c55\u51fa\u4f86\u7684\u5206\u5b50\u5de5\u5177\uff08Techniques of molecular biology\uff09\uff08\u8a3b2\uff09\u6216\u65b0\u8cea\u9ad4\uff08Plasmid\uff09\u5bc4\u5b58\u65bc\u975e\u71df\u5229\u4e8b\u696d\u7684Addgene \u516c\u53f8\uff0c\u4ee5\u63d0\u4f9b\u5176\u4ed6\u6709\u8208\u8da3\u7684\u79d1\u5b78\u5bb6\u4f7f\u7528\u3002\u81ea\u5f9e2013\u5e74\u521d\u7814\u7a76\u4eba\u54e1\u9996\u5ea6\u767c\u8868\u4ed6\u5011\u80fd\u4ee5 CRISPR-Cas9 \u6280\u8853\u5207\u5272\u4eba\u985e\u7d30\u80de\u5167\u57fa\u56e0\u9ad4\u4efb\u4f55\u4e00\u7279\u5b9a\u4f4d\u7f6e\u5f8c\uff0cAddgene \u516c\u53f8\u7684\u8a62\u554f\u96fb\u8a71\u5c31\u6c92\u505c\u904e\uff0c\u5206\u5b50\u751f\u7269\u5b78\u5bb6\u5011\u90fd\u6025\u65bc\u7372\u5f97\u6b64\u6280\u8853\uff0c\u80fd\u5920\u7c21\u55ae\u4e14\u6e96\u78ba\u5730\u4fee\u6539\u7de8\u8f2f\u5e7e\u4e4e\u6240\u6709\u751f\u7269\u9ad4\u7684\u57fa\u56e0\u9ad4\u3002<\/p>\n

\u76ee\u524d\u771f\u6b63\u9769\u547d\u6027\u7684\u767c\u5c55\u6b63\u5728\u5be6\u9a57\u5ba4\u88cf\u5c55\u958b\uff0cCRISPR \u63d0\u4f9b\u751f\u7269\u5b78\u5bb6\u6240\u9700\u8981\u7684\u4e00\u5927\u7279\u9ede\uff1a\u7279\u5b9a\u660e\u78ba\u6027\uff0c\u53ef\u4ee5\u6e96\u78ba\u5730\u8fa8\u8b58\u9f90\u5927\u7684\u57fa\u56e0\u9ad4\u88f9\u7279 \u5b9a\u7684\u5e8f\u5217\u3002\u800c\u7de8\u8f2fDNA \u53ea\u662f\u5176\u4e2d\u4e00\u9805\u6280\u80fd\uff0c\u79d1\u5b78\u5bb6\u9032\u4e00\u6b65\u5c07\u6b64\u6280\u8853\u505a\u4e0d\u540c\u7684\u6539\u8b8a\uff0c\u53ef\u4ee5\u5c07\u86cb\u767d\u8cea\u9001\u81f3\u7279\u5b9a\u57fa\u56e0\u9ad4\u4f4d\u7f6e\u4f5c\u8abf\u63a7\uff0c\u4ee5\u555f\u52d5\u6216\u505c\u6b62\u7279\u5b9a\u57fa\u56e0\u8868\u73fe\uff0c\u751a\u81f3\u53ef\u4ee5\u8a2d\u8a08\u6574\u500b\u751f\u7269\u5faa\u74b0\uff0c\u6700\u7d42\u4e86\u89e3\u7d30\u80de\u5167\u7684\u7cfb\u7d71\u548c\u75be\u75c5\u7684\u7d30\u7bc0\u3002 \u7562\u7adfCRISPR-Cas9 \u53ea\u662f\u4e00\u9805\u6280\u8853\uff0c\u6211\u5011\u73fe\u5728\u64c1\u6709\u6b64\u4e00\u5f37\u5927\u7684\u5de5\u5177\uff0c\u6700\u7d42\u4ecd\u9700\u56de\u6b78\u89e3\u6c7a\u751f\u7269\u554f\u984c\u672c\u8eab\uff0c\u90a3\u624d\u662f\u6211\u5011\u7684\u771f\u6b63\u76ee\u7684\u3002<\/p>\n

 <\/p>\n

--
\n\u8a3b1\uff1a\u66f4\u6539 DNA \u5e8f\u5217\uff0c\u91cd\u65b0\u505a\u51fa\u7a81\u8b8a\u7684\u86cb\u767d\u8cea\u3002
\n\u8a3b2\uff1a\u7814\u7a76\u5206\u5b50\u751f\u7269\u5b78\u6240\u4f7f\u7528\u7684\u5de5\u5177\u6216\u6280\u8853\uff0c\u4f8b\u5982\u5206\u5b50\u7d14\u5316\u7e41\u6b96<\/a>\uff08Molecular cloning\uff09\u3001\u805a\u5408\u9176\u9023\u9396\u53cd\u61c9<\/a>\uff08Polymerase chain reaction (PCR)\uff09\u3001\u51dd\u81a0\u96fb\u6cf3<\/a>Gel electrophoresis\u3001\u5357\u65b9[\u6c0f]\u5370\u6f2c\u8853<\/a>Southern blotting\u7b49\u7b49\u3002<\/p>\n

\u53c3\u8003\u6587\u737b:<\/strong>
\n1. Nature 531, 156\u2013159 (10 March 2016) doi:10.1038\/531156a
\n2. Qi, L.S., Larson, M.H., Gilbert, L.A., Doudna, J.A., Weissman, J.S., Arkin, A.P., and Lim, W.A. (2013). Repurposing CRISPR as an RNA-guided
\n3. Dow, L.E., Fisher, J., O'Rourke, K.P., Muley, A., Kastenhuber, E.R., Livshits, G., Tschaharganeh, D.F., Socci, N.D., and Lowe, S.W. (2015). Inducible in vivo genome editing with CRISPR-Cas9. Nature biotechnology 33, 390-394.
\n4. Hilton, I.B., D'Ippolito, A.M., Vockley, C.M., Thakore, P.I., Crawford, G.E., Reddy, T.E., and Gersbach, C.A. (2015). Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers. Nature biotechnology 33, 510-517.
\n5. platform for sequence-specific control of gene expression. Cell 152, 1173-1183.
\n6. Rajagopal, N., Srinivasan, S., Kooshesh, K., Guo, Y., Edwards, M.D., Banerjee, B., Syed, T., Emons, B.J., Gifford, D.K., and Sherwood, R.I. (2016). High-throughput mapping of regulatory DNA. Nature biotechnology 34, 167-174.
\n7. Zetsche, B., Gootenberg, J.S., Abudayyeh, O.O., Slaymaker, I.M., Makarova, K.S., Essletzbichler, P., Volz, S.E., Joung, J., van der Oost, J., Regev, A., et al. (2015). Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell 163, 759-771.
\n8. Polstein, L.R., and Gersbach, C.A. (2015). A light-inducible CRISPR-Cas9 system for control of endogenous gene activation. Nature chemical biology 11, 198-200.<\/p>\n

 <\/p>\n

\"\u52a0\u5165\u597d\u53cb\"<\/a><\/p>\n","protected":false},"excerpt":{"rendered":"

\u5c0d\u5206\u5b50\u751f\u7269\u5b78\u5bb6\u800c\u8a00\uff0cCRISPR \u662f\u4e00\u975e\u5e38\u5f37\u5927\u7684\u5de5\u5177\uff0c\u53ef\u4ee5\u7528\u4f86\u66f4\u77ad\u89e3\u57fa\u56e0\u9ad4\u5982\u4f55\u904b\u4f5c\uff0c\u4ee5\u53ca\u63a2\u8a0e\u66f4\u6df1\u5165\u7684\u554f\u984c\u30022016\u5e74\u300c\u81ea\u7136\u300d\u671f\u520a\uff08Nature \uff09\u6574\u7406\u51fa CRISPR-Cas9 \u767c\u5c55\u7684\u4e94\u500b\u65b9\u5411\uff0c\u53ca\u5176\u5982\u4f55\u6539\u8b8a\u79d1\u5b78\u5bb6\u5011\u7814\u7a76\u7684\u65b9\u6cd5\u3002<\/p>\n","protected":false},"author":20,"featured_media":26917,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[3293,1025],"tags":[3109,3075,3108,3148,129],"aioseo_notices":[],"jetpack_featured_media_url":"https:\/\/case.ntu.edu.tw\/blog\/wp-content\/uploads\/2017\/02\/4972914961_5b8c976bd1_b-1024x640.jpg","_links":{"self":[{"href":"https:\/\/case.ntu.edu.tw\/blog\/index.php?rest_route=\/wp\/v2\/posts\/26830"}],"collection":[{"href":"https:\/\/case.ntu.edu.tw\/blog\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/case.ntu.edu.tw\/blog\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/case.ntu.edu.tw\/blog\/index.php?rest_route=\/wp\/v2\/users\/20"}],"replies":[{"embeddable":true,"href":"https:\/\/case.ntu.edu.tw\/blog\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=26830"}],"version-history":[{"count":4,"href":"https:\/\/case.ntu.edu.tw\/blog\/index.php?rest_route=\/wp\/v2\/posts\/26830\/revisions"}],"predecessor-version":[{"id":31664,"href":"https:\/\/case.ntu.edu.tw\/blog\/index.php?rest_route=\/wp\/v2\/posts\/26830\/revisions\/31664"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/case.ntu.edu.tw\/blog\/index.php?rest_route=\/wp\/v2\/media\/26917"}],"wp:attachment":[{"href":"https:\/\/case.ntu.edu.tw\/blog\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=26830"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/case.ntu.edu.tw\/blog\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=26830"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/case.ntu.edu.tw\/blog\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=26830"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}