{"id":10637,"date":"2017-06-12T13:49:08","date_gmt":"2017-06-12T05:49:08","guid":{"rendered":"http:\/\/biopioneer.com.tw\/?p=10637"},"modified":"2022-07-22T10:20:03","modified_gmt":"2022-07-22T02:20:03","slug":"b3galt5-crispr-%e7%b4%b0%e8%83%9e%e6%a0%aab3galt5-crispr-knock-out-mcf7-cell-line-%e8%b2%a8%e8%99%9fk0163157","status":"publish","type":"post","link":"http:\/\/biopioneer.com.tw\/?p=10637","title":{"rendered":"B3GALT5 CRISPR \u7d30\u80de\u682a|B3GALT5 CRISPR Knock Out MCF7 Cell Line |\u8ca8\u865fK0163157"},"content":{"rendered":"

B3GALT5 CRISPR Knock Out MCF7 Cell Line (Human)<\/strong> \u7d30\u80de\u57f9\u990a\u689d\u4ef6<\/strong> |<\/strong>\u8ca8\u865f<\/strong>K0163157<\/strong><\/p>\n

CRISPR\u7d30\u80de\u8cc7\u6599: B3GALT5 Accession Number NM_033173<\/a> | MCF7<\/p>\n

\"\"<\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
Vector<\/td>\npLenti-U6-sgRNA-PGK-Neo<\/a><\/td>\n<\/tr>\n
Target Sequence<\/td>\nSequence available upon order placement<\/td>\n<\/tr>\n
Format<\/td>\nCell line<\/td>\n<\/tr>\n
Parental Cell Line<\/td>\nMCF7<\/td>\n<\/tr>\n
Drug resistance<\/td>\nPuromycin (0.5\u03bcg\/ml)<\/td>\n<\/tr>\n
Culture Condition<\/td>\nCulture Condition The base medium for this cell line is DMEM, 10%FBS and 1%P\/S, 37\u00b0C, 5% CO2<\/sub><\/td>\n<\/tr>\n
Appearance<\/td>\nLiquid<\/td>\n<\/tr>\n
Storage<\/td>\n-130\u00b0C<\/td>\n<\/tr>\n
Shipping<\/td>\nShipped with dry ice<\/td>\n<\/tr>\n
Additional Materials<\/td>\n– CRISPR Genomic Cleavage Detection Kit (G932)<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

\u57fa\u56e0\u7d44\u5de5\u7a0b\u5de5\u5177<\/strong><\/p>\n

\u5728\u73fe\u4ee3\u7cfb\u7d71\u4e2d\uff0c\u4f7f\u7528CRISPR Cas9\u6280\u8853\u7684\u9776\u5411\u57fa\u56e0\u7d44\u7de8\u8f2f\u6709\u5169\u500b\u7d44\u6210\u90e8\u5206\uff1a1\uff09\u5167\u5207\u6838\u9178\u9176;\u548c2\uff09\u77ed\u5c0e\u5411RNA\uff08\u57162\uff09\u3002\u6838\u9178\u5167\u5207\u9176\u662f\u4f86\u81ea\u5316\u81bf\u6027\u93c8\u7403\u83cc\u7684\u7d30\u83ccCas9\u6838\u9178\u9176\u86cb\u767d\u3002 Cas9\u6838\u9178\u9176\u5177\u6709\u5207\u5272\u96d9\u93c8DNA\u7684\u5169\u500bDNA\u5207\u5272\u7d50\u69cb\u57df\uff08RuvC1\u548cHNH\u6a23\u6838\u9178\u9176\u7d50\u69cb\u57df\uff09\uff0c\u7522\u751f\u96d9\u93c8\u65b7\u88c2\uff08DSB\uff09\uff084\uff09\u3002 gRNA\u662f\u5de5\u7a0b\u5316\u7684\u55ae\u93c8\u5d4c\u5408RNA\uff0c\u5c07\u7d30\u83cctracrRNA\u7684\u652f\u67b6\u529f\u80fd\u8207\u7d30\u83cccrRNA\u7684\u7279\u7570\u6027\u7d50\u5408\uff086\uff09\u3002 gRNA\u76845’\u672b\u7aef\u7684\u6700\u5f8c\u4e00\u500b20bp\u4f5c\u70ba\u6b78\u5de2\u88dd\u7f6e\uff0c\u901a\u904eRNA-DNA\u9e7c\u57fa\u914d\u5c0d\uff0c\u5c07Cas9 \/ gRNA\u8907\u5408\u7269\u52df\u96c6\u5230\u539f\u59cb\u76f8\u9130\u57fa\u5e8f\uff08PAM\uff09\u4e0a\u6e38\u7684\u7279\u5b9aDNA\u9776\u4f4d\u9ede\u3002\u4e0d\u540c\u83cc\u682a\u548cCRISPR Cas\u86cb\u767d\u8cea\u985e\u578b\u7684PAM\u5e8f\u5217\u4e0d\u540c\uff0c\u5316\u81bf\u6027\u93c8\u7403\u83ccCas9\u7684\u5e8f\u5217\u662f5′-NGG\uff08\u53c3\u898bProtospacer Adjacent Motif\uff09\u3002\u56e0\u6b64\uff0c\u4eca\u5929\u53ef\u7528\u7684\u9069\u61c9CRISPR Cas9\u7cfb\u7d71\u53ef\u4ee5\u91dd\u5c0d\u4efb\u4f555′-N20-NGG DNA\u5e8f\u5217\u4e26\u7522\u751f\u7cbe\u78ba\u7684\u96d9\u93c8\u65b7\u88c2\uff087\uff09\u3002\u7136\u5f8c\u901a\u904e\u5728\u5e7e\u4e4e\u6240\u6709\u7d30\u80de\u985e\u578b\u548c\u751f\u7269\u9ad4\u4e2d\u767c\u73fe\u7684\u5169\u7a2e\u901a\u7528\u4fee\u5fa9\u6a5f\u5236\u4e4b\u4e00\u4f86\u4fee\u5fa9DSB\uff1a\u975e\u540c\u6e90\u672b\u7aef\u9023\u63a5\uff08NHEJ\uff09\u6216\u540c\u6e90\u6027\u5b9a\u5411\u4fee\u5fa9\uff08HDR\uff09\u3002\"\"<\/p>\n

\u975e\u540c\u6e90\u672b\u7aef\u9023\u63a5\uff08NHEJ\uff09\u4fee\u5fa9\u9014\u5f91<\/strong><\/p>\n

\u5728\u6c92\u6709\u5408\u9069\u7684\u4fee\u5fa9\u6a21\u677f\u7684\u60c5\u6cc1\u4e0b\uff0cNHEJ\u4fee\u5fa9\u9014\u5f91\u662f\u7528\u65bc\u4fee\u5fa9\u96d9\u93c8\u65b7\u88c2\u7684\u6613\u932f\u4fee\u5fa9\u6a5f\u5236\u3002 NHEJ\u9014\u5f91\u5617\u8a66\u5c07DSB\u7684\u5207\u5272\u672b\u7aef\u9023\u63a5\u5728\u4e00\u8d77\u3002 \u7136\u800c\uff0c\u8a72\u904e\u7a0b\u901a\u5e38\u5c0e\u81f4DSB\u4f4d\u9ede\u7684\u63d2\u5165\u6216\u7f3a\u5931\uff08InDels\uff09\u7a81\u8b8a\uff0c\u5c0e\u81f4\u5e40\u4f4d\u79fb\u6216\u5f15\u5165\u6c38\u4e45\u6027\u7834\u58de\u9776\u57fa\u56e0\u958b\u653e\u95b1\u8b80\u6846\u67b6\u7684\u6210\u719f\u524d\u7d42\u6b62\u5bc6\u78bc\u5b50\uff08\u57163\uff09\u3002 \u96d6\u7136NHEJ\u7684InDel\u7d50\u679c\u4ecd\u7136\u5f88\u5927\u7a0b\u5ea6\u4e0a\u662f\u96a8\u6a5f\u7684\uff0c\u4f46\u79d1\u5b78\u5bb6\u5011\u53ef\u4ee5\u901a\u904e\u5c07\u76ee\u6a19\u57fa\u56e0\u7684gRNA\u9776\u5411\u76ee\u6a19\u57fa\u56e0\u7684N\u672b\u7aef\u4f86\u78ba\u4fdd\u6700\u5927\u7684\u57fa\u56e0\u7834\u58de\u3002 \u9019\u5c07\u78ba\u4fdd\u5e40\u8f49\u79fb\u7a81\u8b8a\u4e0d\u5c0e\u81f4\u90e8\u5206\u529f\u80fd\u6027\u57fa\u56e0\u7522\u7269\u3002 \u8a2d\u8a08\u7b2c\u4e00\u500b\u6216\u7b2c\u4e8c\u500b\u5916\u986f\u5b50\u4e2d\u7684gRNA\u662f\u597d\u7684\u505a\u6cd5\uff0c\u4e26\u4e14\u5728\u5229\u7528NHEJ\u4fee\u5fa9\u9014\u5f91\u6642\u907f\u514d\u9776\u5411\u57fa\u56e0\u7684\u5167\u542b\u5b50\u5340\u57df\uff087\uff09\u3002\"\"<\/p>\n

\u8207Cas9\u6838\u9178\u9176\u7684\u540c\u6e90\u6027\u5b9a\u5411\u4fee\u5fa9\uff08HDR\uff09<\/strong><\/p>\n

\u9664\u4e86NHEJ\uff0c\u7d30\u80de\u80fd\u5920\u5229\u7528\u88ab\u7a31\u70ba\u540c\u6e90\u6027\u5b9a\u5411\u4fee\u5fa9\uff08HDR\uff09\u9014\u5f91\u7684\u66f4\u7cbe\u78ba\u7684\u4fee\u5fa9\u6a5f\u5236\u3002 \u9019\u7a2e\u4fee\u5fa9\u6a5f\u5236\u53ef\u4ee5\u7528\u65bc\u5f15\u5165\u5c0d\u57fa\u56e0\u7d44DNA\u7684\u7279\u7570\u6027\u6838\u82f7\u9178\u4fee\u98fe\uff088\uff09\u3002 \u5728\u8a72\u65b9\u6cd5\u4e2d\uff0c\u8207\u9069\u7576\u7684gRNA\u548cCas9\u6838\u9178\u9176\u4e00\u8d77\u5c07\u8207\u9810\u671f\u7de8\u8f2f\u90e8\u4f4d\u4e0a\u6e38\u548c\u4e0b\u6e38\u7684\u5e8f\u5217\u5177\u6709\u9ad8\u5ea6\u540c\u6e90\u6027\u7684DNA\u4fee\u5fa9\u6a21\u677f\u5f15\u5165\u7d30\u80de\u3002 \u5728\u9019\u7a2e\u5408\u9069\u7684\u6a21\u677f\u7684\u5b58\u5728\u4e0b\uff0c\u8f03\u4e0d\u6613\u51fa\u932f\u7684HDR\u6a5f\u5236\u53ef\u4ee5\u5fe0\u5be6\u5730\u901a\u904e\u91cd\u7d44\u5c0dCas9\u8a98\u5c0e\u7684DSB\u4f4d\u9ede\u9032\u884c\u6240\u9700\u7684\u6539\u8b8a\uff08\u57164\uff09\u3002 \u5728\u8a2d\u8a08\u4fee\u5fa9\u6a21\u677f\u6642\uff0c\u8acb\u78ba\u4fdd\u76ee\u6a19\u5e8f\u5217\u4e0d\u7dca\u96a8\u8457PAM\u5e8f\u5217\u6216PAM\u5e8f\u5217\u88ab\u6392\u9664\u6216\u7a81\u8b8a\u3002 \u9019\u662f\u70ba\u4e86\u907f\u514d\u76f8\u540c\u7684CRISPR Cas9\u7cfb\u7d71\u4fee\u5fa9\u6a21\u677f\u7684\u52a3\u5316\u3002\"\"<\/p>\n

Protospacer<\/strong>\u76f8\u9130\u4e3b\u984c<\/strong><\/p>\n

CRISPR Cas9\u7cfb\u7d71\u53ef\u4ee5\u901a\u904egRNA\u7684\u8a2d\u8a08\u9776\u5411\u4efb\u4f55\u57fa\u56e0\u7d44\u5340\u57df\uff0c\u7136\u800c\uff0c\u7cfb\u7d71\u7684\u7279\u7570\u6027\u53d6\u6c7a\u65bc\u4f4d\u65bc\u9776\u5e8f\u5217\u7dca\u9130\u4e0b\u6e38\u7684\u555f\u52d5\u5b50\u76f8\u9130\u5e8f\u5217\uff08PAM\uff09\uff08\u57165\uff09\uff083\uff09\u3002 \u4f5c\u70ba\u7d30\u83cc\u548c\u53e4\u7d30\u83cc\u514d\u75ab\u7cfb\u7d71\uff0cPAM\u8b58\u5225\u5e8f\u5217\u6fc0\u6d3bCas9\u7684\u6838\u9178\u9176\u7d50\u69cb\u57df\uff0c\u5f9e\u800c\u4f5c\u70ba\u5340\u5206\u81ea\u8eab\u8207\u975e\u81ea\u8eab\u7684\u624b\u6bb5\uff08\u5373\u9632\u6b62CRISPR\u57fa\u56e0\u5ea7\u88ab\u9776\u5411\uff09\uff087\uff09\u3002<\/p>\n

\"\"<\/p>\n

PAM\u8b58\u5225\u5e8f\u5217\u53d6\u6c7a\u65bc\u884d\u751fCas9\u6838\u9178\u9176\u7684\u7d30\u83cc\u7684\u7a2e\u985e\u548c\u985e\u578b\u3002 \u6700\u5e38\u7528\u7684II\u578bCRISPR\u7cfb\u7d71\u4f7f\u7528\u4f86\u81ea\u5316\u81bf\u6027\u93c8\u7403\u83cc\u7684Cas9\u6838\u9178\u9176\u3002 \u9019\u7a2e\u7279\u7570\u7684\u6838\u9178\u9176\u5728gRNA\u5e8f\u5217\u76843’\u672b\u7aef\u8b58\u52255′-NGG\u3002 \u5176\u4ed6\u5e02\u552e\u7684Cas9\u53ef\u4ee5\u8b58\u5225\u5176\u4ed6PAM\u5e8f\u5217\uff08\u88681\uff09\u3002<\/p>\n

Table 1:<\/strong> PAM Sequences from Different Species and Subtypes of Cas Nucleases<\/p>\n

\n
\n
\n
\n
\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Species<\/th>\nSubtype<\/th>\nPAM Sequence<\/th>\n<\/tr>\n<\/tbody>\n
S. pyogenes<\/em><\/td>\nII<\/td>\n5′-NGG<\/td>\n<\/tr>\n
S. solfataricus<\/em><\/td>\nI-A1<\/td>\n5′-CCN<\/td>\n<\/tr>\n
S. solfataricus<\/em><\/td>\nI-A2<\/td>\n5′-TCN<\/td>\n<\/tr>\n
H. walsbyi<\/em><\/td>\nI-B<\/td>\n5′-TTC<\/td>\n<\/tr>\n
E. coli<\/em><\/td>\nI-E<\/td>\n5′-AWG<\/td>\n<\/tr>\n
E. coli<\/em><\/td>\nI-F<\/td>\n5′-CC<\/td>\n<\/tr>\n
P. aeruginosa<\/em><\/td>\nI-F<\/td>\n5′-CC<\/td>\n<\/tr>\n
S. Thermophilus<\/em><\/td>\nII-A<\/td>\n5′-NNAGAA<\/td>\n<\/tr>\n
S. agelactiae<\/em><\/td>\nII-A<\/td>\n5′-NGG<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n
\n
\n
\n
\n
\n
\u00a0Off-Target Mutations<\/strong><\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n

\u4f7f\u7528CRISPR Cas9\u4f5c\u70ba\u57fa\u56e0\u7d44\u7de8\u8f2f\u5de5\u5177\u6642\u7684\u4e00\u500b\u91cd\u8981\u8003\u616e\u662f\u812b\u9776\u88c2\u89e3\u767c\u751f\u7684\u7a0b\u5ea6\u3002\u96e2\u9776\u4e8b\u4ef6\u53ef\u80fd\u6703\u5c0e\u81f4InDel\u7a81\u8b8a\u4e0d\u662f\u6700\u521d\u7684\u610f\u5716\uff0c\u56e0\u6b64\u6703\u640d\u5bb3\u6240\u7372\u5f97\u7684\u8868\u578b\u7d50\u679c\u3002\u5e7e\u9805\u7814\u7a76\u5df2\u7d93\u8a55\u4f30\u4e86CRISPR Cas9\u7cfb\u7d71\u7684\u7279\u7570\u6027\uff0c\u4e26\u4e14\u5df2\u7d93\u8868\u660e\u901a\u5e38\u53ef\u4ee5\u8010\u53d7\u8207gRNA\u768420\u500b\u9e7c\u57fa\u5c0d\u9776\u5411\u5340\u76845’\u672b\u7aef\u7684\u932f\u914d\u3002 \uff086\uff09\uff089\uff09\uff0810\uff09\uff0811\uff09\uff0812\uff09\u7136\u800c\uff0c\u5f88\u96e3\u9810\u6e2c\u9019\u4e9b\u5931\u914d\u5982\u4f55\u5f71\u97ffCRISPR Cas9\u7cfb\u7d71\u7684\u975e\u76ee\u6a19\u6548\u61c9\u3002\u5df2\u7d93\u6709\u4e00\u4e9b\u60c5\u6cc1\u5831\u5c0e\u4e86\u5728gRNA\u7684\u9776\u5411\u5340\u57df\u76845’\u672b\u7aef\u7684\u4e0d\u5339\u914d\u4e0d\u88ab\u8010\u53d7\u7684\u60c5\u6cc1\uff0c\u4ee5\u53ca\u5141\u8a31\u9776\u5411\u5340\u57df3’\u7aef\u7684\u932f\u914d\u7684\u5176\u4ed6\u60c5\u6cc1\uff0813\uff09\u3002\u7e3d\u9ad4\u4f86\u8aaa\uff0cCRISPR Cas9\u7cfb\u7d71\u5f15\u5165\u7684\u975e\u76ee\u6a19\u6548\u61c9\u5728\u983b\u7387\u4e0a\u662f\u8b8a\u5316\u7684\uff0c\u4e26\u4e14\u5177\u6709\u6311\u6230\u6027\u4f86\u9810\u6e2c\uff087\uff09\u3002<\/p>\n

\u53ef\u4ee5\u4f7f\u7528\u5e7e\u7a2e\u53ef\u7528\u7684\u65b9\u6cd5\u4f86\u6700\u5c0f\u5316\u9019\u4e9b\u812b\u9776\u6548\u61c9\u3002\u901a\u904e\u5c07gRNA\u5e8f\u5217\u4ed4\u7d30\u8a2d\u8a08\u5230\u76ee\u6a19\u57fa\u56e0\uff0c\u53ef\u4ee5\u6700\u5927\u9650\u5ea6\u5730\u6e1b\u5c11\u76ee\u6a19\u6548\u61c9\uff08\u53c3\u898b\u8a2d\u8a08gRNA\u5e8f\u5217\u7684\u6307\u5c0e\uff09\u3002\u901a\u5e38\u61c9\u7528\u7684\u53e6\u4e00\u7a2e\u65b9\u6cd5\u662f\u4f7f\u7528Cas9\u6838\u9178\u5167\u5207\u9176\u7684\u8b8a\u9ad4\u5f62\u5f0f\u3002\u8a72\u8b8a\u9ad4\u5177\u6709\u4f7f\u5f97\u91ce\u751f\u578bCas9\uff08Cas9Noase\uff09\u7684\u5207\u5272\u7d50\u69cb\u57df\u4e2d\u7684\u4e00\u500b\u5931\u6d3b\u7684\u7a81\u8b8a\uff0c\u4e26\u5c07\u5728\u7a0d\u5f8c\u8a73\u7d30\u8a0e\u8ad6\u3002\u6700\u7d42\u7684\u65b9\u6cd5\u662f\u6e1b\u5c11gRNA\u768420\u500b\u9e7c\u57fa\u5c0d\u9776\u5411\u5e8f\u5217\u3002\u5df2\u7d93\u986f\u793a\u9019\u4e9b\u622a\u77ed\u7684gRNA\uff08tru-gRNA\uff09\u5728\u5c07Cas9\u6838\u9178\u9176\u5f15\u5c0e\u5230\u9810\u5b9a\u9776\u4f4d\u9ede\u6642\u8207\u5168\u9577gRNA\u4e00\u6a23\u6709\u6548\u3002 Tru-gRNA\u5728\u812b\u9776\u5e8f\u5217\u4e0a\u8868\u73fe\u51fa\u964d\u4f4e\u7684\u8a98\u8b8a\u6548\u61c9\uff0c\u4e26\u4e14\u5c0d\u55ae\u4e00\u6216\u96d9\u91cd\u932f\u914d\u66f4\u654f\u611f\u3002 \uff0814\uff09<\/p>\n

Cas9<\/strong>\u96d9\u7a81\u8b8a\u9ad4<\/strong><\/p>\n

Cas9\u96d9\u7a81\u8b8a\u9ad4\u6216Null\u7a81\u8b8a\u9ad4\u662f\u901a\u904e\u7a81\u8b8a\u91ce\u751f\u578bCas9\u7684\u5169\u500b\u5207\u5272\u7d50\u69cb\u57df\u800c\u7522\u751f\u7684\u3002 \u9019\u6a23\u7684Cas9\u86cb\u767d\u4fdd\u7559\u5176\u901a\u904egRNA\u8207\u57fa\u56e0\u7d44DNA\u7d50\u5408\u7684\u80fd\u529b\uff1a\u57fa\u56e0\u7d44DNA\u9e7c\u57fa\u914d\u5c0d\u3002 \u8207Cas9 Nuclease\u548cCas9Noase\u53ef\u4ee5\u5be6\u73fe\u7684\u6c38\u4e45\u57fa\u56e0\u7834\u58de\u4e0d\u540c\uff0cCas9 Null\u7a81\u8b8a\u9ad4\u4e0d\u5f15\u5165\u4efb\u4f55\u57fa\u56e0\u7d44\u4fee\u98fe\u3002 \u901a\u904e\u5c07Cas9\u96d9\u7a81\u8b8a\u9ad4\u8207\u5176\u4ed6\u6548\u61c9\u86cb\u767d\u878d\u5408\uff0cCRISPR Cas9\u7cfb\u7d71\u53ef\u4ee5\u64f4\u5c55\u5176\u57fa\u56e0\u8abf\u63a7\uff0c\u57fa\u56e0\u7d44\u6210\u50cf\uff0c\u67d3\u8272\u8cea\u6216DNA\u4fee\u98fe\u548c\u67d3\u8272\u8cea\u514d\u75ab\u6c89\u6fb1\u7684\u4f5c\u7528\uff08\u57168\uff09\uff0815\uff09\uff0816\uff09\uff0817\uff09\uff0818\uff09\uff0819\uff09\u3002<\/p>\n

\u8207\u86cb\u767d\u8cea\u57fa\u56e0\u7d44\u7de8\u8f2f\u5de5\u5177\u7684\u6bd4\u8f03<\/strong><\/p>\n

\u6709\u5169\u7a2e\u5e38\u898b\u7684\u57fa\u65bc\u86cb\u767d\u8cea\u7684\u57fa\u56e0\u7d44\u7de8\u8f2f\u5de5\u5177\uff1a1\uff09\u92c5\u6307\u6838\u9178\u9176\uff08ZFN\uff09\u548c2\uff09\u8f49\u9304\u6fc0\u6d3b\u5b50\u6a23\u6548\u61c9\u6838\u9178\u9176\uff08TALENs\uff09\u3002 \u88683\u6bd4\u8f03\u548c\u5c0d\u6bd4\u5169\u7a2e\u57fa\u65bc\u86cb\u767d\u8cea\u7684\u65b9\u6cd5\u8207\u57fa\u65bcRNA\u7684CRISPR Cas9\u7cfb\u7d71\uff0820\uff09\u3002<\/p>\n

Table 3:<\/strong> Comparison of current genome editing technologies<\/p>\n\n\n\n\n\n\n\n\n\n
<\/td>\nZFNs<\/strong><\/td>\nTALENs<\/strong><\/td>\nCRISPR Cas9<\/strong><\/td>\n<\/tr>\n
DNA Binding Domain<\/td>\nCys2-His2 DNA Binding Protein<\/td>\nConserved Amino Acid Repeated Motif<\/td>\nSingle Stranded gRNA<\/td>\n<\/tr>\n
DNA Cleavage Domain<\/td>\nFokI Restriction Endonuclease<\/td>\nFokI Restriction Endonuclease<\/td>\nCas9 Endonuclease<\/td>\n<\/tr>\n
Guiding Mechanism<\/td>\nProtein Guided<\/td>\nProtein Guided<\/td>\nRNA Guided<\/td>\n<\/tr>\n
Ease of Design<\/td>\n*<\/td>\n***<\/td>\n*****<\/td>\n<\/tr>\n
Minimized off-target Effects<\/td>\n*****<\/td>\n*****<\/td>\n***<\/td>\n<\/tr>\n
Multiplexibility<\/td>\n**<\/td>\n**<\/td>\n*****<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u57fa\u56e0\u6cbb\u7642<\/strong><\/p>\n

2012\u5e74\uff0cCRISPR Cas9\u9996\u6b21\u88ab\u8b49\u660e\u662f\u4eba\u985e\u7d30\u80de\u57f9\u990a\u57fa\u56e0\u7d44\u7de8\u8f2f\u5de5\u5177\uff0821\uff09\u3002 \u6b64\u5f8c\uff0c\u5b83\u5df2\u88ab\u5ee3\u6cdb\u7528\u65bc\u751f\u7269\u9ad4\uff0c\u5305\u62ec\u9eb5\u5305\u9175\u6bcd\uff0822\uff09\uff0c\u690d\u7269\uff0823\uff09\uff0c\u6591\u99ac\u9b5a\uff0824\uff09\uff0c\u679c\u8805\uff0825\uff09\uff0c\u7dda\u87f2\uff0826\uff09\uff0c\u5c0f\u9f20\uff0827\uff09\u548c\u5176\u4ed6\u5e7e\u7a2e\u751f\u7269\u9ad4\u3002CRISPR Cas9\u5177\u6709\u6210\u70ba\u6cbb\u7642\u907a\u50b3\u75be\u75c5\u548c\u75c5\u6bd2\u611f\u67d3\u7684\u6cbb\u7642\u5291\u7684\u6f5b\u529b\u3002 \u4e00\u500b\u7814\u7a76\u7d44\u4f7f\u7528gRNA\u9776\u5411HIV-1\u57fa\u56e0\u7d44\u7684\u9577\u672b\u7aef\u91cd\u8907\u555f\u52d5\u5b50\uff0c\u4ee5\u986f\u8457\u6291\u5236\u5176\u5728\u611f\u67d3\u7684\u4eba\u985e\u7d30\u80de\u4e2d\u7684\u8868\u9054\uff0828\uff09\u3002 \u96a8\u8457\u7814\u7a76\u7684\u5feb\u901f\u767c\u5c55\uff0c\u4f7f\u7528\u548c\u6539\u9032\u4e86CRISPR Cas9\u7cfb\u7d71\uff0c\u76f4\u5230\u63d0\u4f9b\u53ef\u884c\u7684\u57fa\u56e0\u6cbb\u7642\u5291\u624d\u6703\u9577\u4e45\u3002 \u4e8b\u5be6\u4e0a\uff0c\u5df2\u7d93\u5efa\u7acb\u4e86\u8af8\u5982Editas Medicine\u548cCRISPR Therapeutics\u7b49\u516c\u53f8\uff0c\u76ee\u7684\u662f\u4f7f\u7528CRISPR Cas9\u7cfb\u7d71\u958b\u767c\u57fa\u56e0\u6cbb\u7642\u3002<\/p>\n

\u76f8\u95dc\u5546\u54c1<\/strong> CRISPR\u6aa2\u6e2c\u5957\u7d44<\/a><\/strong> | CRISPR\u57fa\u56e0\u5254\u9664\u7d30\u80de\u682a<\/a><\/strong> | CASPASE9 (D10A) NLS<\/a><\/strong><\/p>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n","protected":false},"excerpt":{"rendered":"

B3GALT5 CRISPR Knock Out MCF7 Cell Line (Human) \u7d30\u80de\u57f9\u990a\u689d\u4ef6 […]<\/p>\n","protected":false},"author":3,"featured_media":10635,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[458],"tags":[3655],"_links":{"self":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/10637"}],"collection":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=10637"}],"version-history":[{"count":2,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/10637\/revisions"}],"predecessor-version":[{"id":10639,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/10637\/revisions\/10639"}],"wp:featuredmedia":[{"embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/media\/10635"}],"wp:attachment":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=10637"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=10637"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=10637"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}