{"id":9660,"date":"2025-03-24T22:21:58","date_gmt":"2025-03-24T14:21:58","guid":{"rendered":"https:\/\/fanyiming.life\/?p=9660"},"modified":"2025-12-21T01:03:52","modified_gmt":"2025-12-20T17:03:52","slug":"9660","status":"publish","type":"post","link":"https:\/\/fanyiming.life\/?p=9660","title":{"rendered":"\u7528\u4e8espCas9 KO\u7684sgRNA\u8bbe\u8ba1"},"content":{"rendered":"\n

\u7528\u4e8espCas9 KO\u7684sgRNA\u8bbe\u8ba1\u539f\u5219\uff1a<\/p>\n\n\n\n

    \n
  1. \u5bf9\u4e8e\u6709\u591a\u4e2asplicing variants\u7684\u57fa\u56e0\uff0csgRNA\u5e94\u8be5\u8bbe\u8ba1\u5728\u6240\u6709variants\u7684\u5171\u6709\u533a\u6bb5\uff08NCBI\u53ef\u4ee5\u770b\uff09\uff1b<\/li>\n\n\n\n
  2. \u5728\u5916\u663e\u5b50\u800c\u975e\u5185\u542b\u5b50\u4e0a\u8bbe\u8ba1sgRNA\uff1b<\/li>\n\n\n\n
  3. \u5bf9\u4e8emRNA\uff0c\u5728CDS\u800c\u975e5’\/3’UTR\u4e0a\u8bbe\u8ba1sgRNA\uff1b<\/li>\n\n\n\n
  4. \u5bf9\u4e8emRNA\uff0c\u5728CDS\u7684\u80f8\u90e8\u8bbe\u8ba1sgRNA\uff0c\u4e0d\u5728\u5934\u90e8\uff08\u9632\u6b62\u4e0b\u6e38alternative AUG\u8d77\u59cb\u7ffb\u8bd1\uff09\u4e5f\u4e0d\u5728\u540e\u534a\u6bb5\uff08\u9632\u6b62\u4ea7\u751f\u4fdd\u7559\u90e8\u5206\u529f\u80fd\u7684RNA\uff09\uff1b<\/li>\n\n\n\n
  5. \u5bf9\u4e8e\u4efb\u4f55\u57fa\u56e0\uff0c\u5e94\u8be5\u8bbe\u8ba1\u591a\u6761sgRNAs\uff0c\u9632\u6b62\u5b9e\u9a8c\u73b0\u8c61\u7531\u8131\u9776\u6548\u5e94\u4ea7\u751f\u3002<\/li>\n<\/ol>\n\n\n\n
    \n

    \u5f20\u6d9b\u8bf4Alternative AUG\u8d77\u59cb\u7684\u53ef\u80fd\u6bd4\u8f83\u5c0f\uff0c\u5927\u591a\u6570\u65f6\u5019\u8fd8\u662f\u9760\u4e0a\u6e38\u597d\u4e00\u70b9\u3002<\/p>\n\n\n\n

    \u53e6\u5916\uff0c\u6709\u65f6\u5019RefSeq\u6709\u5f88\u591a\u6761RNA variant\uff0c\u5e76\u4e0d\u4e00\u5b9a\u4f60\u6240\u7528\u5b9e\u9a8c\u6750\u6599\u5185\u5168\u90fd\u6709\uff0c\u5927\u90e8\u5206\u60c5\u51b5\u8fd8\u662f\u5c311~2\u6761\u5360\u7edd\u5bf9\u4f18\u52bf\uff0c\u53ef\u4ee5\u7ed3\u5408\u76f8\u5e94\u7ec6\u80de\u7684RNA seq\u6570\u636e\u68c0\u67e5\u3002\u8fd8\u53ef\u4ee5\u901a\u8fc7\u70b9\u5f00\u67d0\u4e00\u6761RefSeq\uff0c\u770bNCBI\u6761\u76ee\u91cc\u7684\u53c2\u8003\u6587\u732e\u591a\u4e0d\u591a\uff0c\u5982\u679c\u591a\u7684\u8bdd\u8bf4\u660e\u53ef\u80fd\u662f\u4e3b\u8981\u7684\u8f6c\u5f55\u672c\u3002<\/p>\n<\/blockquote>\n\n\n\n

    Uniprot\u91cc\u67d0\u4e2a\u57fa\u56e0\u7684\u201csequence\u201d\u680f\u7684\u5e8f\u5217\u4e5f\u53ef\u53c2\u8003\uff1a<\/p>\n\n\n\n

    \n

    \u5bf9\u4e8e\u4eba\u7c7b\u57fa\u56e0\uff0c\u5f53\u68c0\u7d22\u67d0\u4e00\u4e2a\u5177\u4f53\u7684\u57fa\u56e0\u65f6\uff0c\u7ed3\u679c\u9875\u9762\u7684Sequence\u533a\u57df\u901a\u5e38\u4f1a\u9009\u5b9a\u4e00\u4e2a\u7279\u5b9a\u7684\u8f6c\u5f55\u672c\u7f16\u7801\u7684\u86cb\u767d\u8d28\uff08isoform\uff09\u4f5c\u4e3a\u201ccanonical\u201d\u5e8f\u5217\u3002\u8fd9\u4e00\u5e8f\u5217\u4e0eNCBI RefSeq\u6570\u636e\u5e93\u4e2d\u7684\u67d0\u4e00\u4e2a\u8f6c\u5f55\u672c\u5b8c\u5168\u5bf9\u5e94\uff0c\u4f53\u73b0\u4e86\u5176\u6743\u5a01\u6027\u548c\u6807\u51c6\u6027\u3002<\/p>\n\n\n\n

    UniProt\u5728\u786e\u5b9a\u67d0\u4e2a\u86cb\u767d\u8d28\u7684\u201ccanonical\u201d\u5e8f\u5217\u65f6\uff0c\u9075\u5faa\u7740\u4e00\u5957\u4e25\u683c\u7684\u6807\u51c6\uff0cUniProt\u9009\u62e9\u201ccanonical\u201d\u5e8f\u5217\u5fc5\u987b\u6ee1\u8db3\u4ee5\u4e0b\u56db\u79cd\u6807\u51c6\u4e2d\u7684\u81f3\u5c11\u4e00\u79cd\uff1a<\/p>\n\n\n\n

      \n
    1. \u6700\u666e\u904d<\/li>\n\n\n\n
    2. \u4e0e\u5176\u4ed6\u7269\u79cd\u4e2d\u7684\u540c\u6e90\u5e8f\u5217\u6700\u76f8\u4f3c<\/li>\n\n\n\n
    3. \u57fa\u4e8e\u5176\u957f\u5ea6\u6216\u6c28\u57fa\u9178\u7ec4\u6210\uff0c\u80fd\u591f\u6700\u6e05\u6670\u5730\u63cf\u8ff0\u7ed3\u6784\u57df\u3001\u4e9a\u578b\u3001\u591a\u6001\u6027\u3001\u7ffb\u8bd1\u540e\u4fee\u9970\u7b49<\/li>\n\n\n\n
    4. \u5728\u7f3a\u4e4f\u4efb\u4f55\u4fe1\u606f\u7684\u60c5\u51b5\u4e0b\uff0c\u9009\u62e9\u6700\u957f\u7684\u5e8f\u5217<\/li>\n<\/ol>\nhttps:\/\/mp.weixin.qq.com\/s\/3GaJhdO42ck_tJvAT9qq0Q<\/a><\/cite><\/blockquote>\n\n\n\n

      \u5b9e\u64cd\u4e2d\u6700\u8fc5\u901f\u7684\u65b9\u6cd5\uff1a<\/p>\n\n\n\n

        \n
      1. \u4ece\u5546\u4e1aCas9 CRISPR human 4x Brunello\u7b5b\u9009\u7684\u6587\u5e93sgRNA\u8868\u683c\u4e2d\u627e\u5230\u6240\u9700\u7684\u57fa\u56e0\uff0c\u67094\u6761sgRNA\uff1b<\/li>\n\n\n\n
      2. \u5728Benchling\u770b\u4e00\u4e0b\u5927\u6982\u7684on-score\u548coff-score\u5408\u4e0d\u5408\u9002\uff1b<\/li>\n\n\n\n
      3. \u5728NCBI\u67e5\u4e00\u4e0b\u5bf9splicing variants\u7684\u8986\u76d6\u7387\uff1b<\/li>\n\n\n\n
      4. \u5982\u679c\u5fc5\u8981\uff0c\u518d\u53bbUniprot\u67e5\u67e5main transcript\u662f\u54ea\u4e2a\uff1b\u5982\u679c\u5fc5\u8981\uff0c\u5728\u81ea\u5df1\u5b9e\u9a8c\u6750\u6599\u7684RNA seq\u6570\u636e\u4e2d\u67e5\u67e5\uff1b<\/li>\n\n\n\n
      5. \u786e\u5b9asgRNA\u5e8f\u5217\u540e\uff0c\u6839\u636e\u6240\u9009\u8d28\u7c92\u786e\u5b9a\u5f15\u7269\u8bbe\u8ba1\u7684\u89c4\u5219\uff08\u63a5\u5934\u5e8f\u5217\uff09\uff0c\u5728\u8fd9\u91cc<\/a>\u751f\u6210\u5f15\u7269\u3002<\/li>\n<\/ol>\n\n\n\n

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

        \u8fd9\u7bc7\u7b14\u8bb0\u7cfb\u7edf\u68b3\u7406\u4e86\u4f7f\u7528spCas9\u8fdb\u884c\u57fa\u56e0\u6572\u9664\u65f6sgRNA\u7684\u8bbe\u8ba1\u539f\u5219\u4e0e\u5b9e\u7528\u6d41\u7a0b\uff0c\u6db5\u76d6\u4e86\u4ece\u9776\u70b9\u9009\u62e9\uff08\u5982\u5171\u6709\u533a\u6bb5\u3001CDS\u4e2d\u90e8\uff09\u3001\u8131\u9776\u8003\u91cf\uff0c\u5230\u7ed3\u5408NCBI\u3001Uniprot\u53caRNA-seq\u6570\u636e\u9a8c\u8bc1\u8f6c\u5f55\u672c\uff0c\u5e76\u63d0\u4f9b\u4e86\u57fa\u4e8e\u6210\u719f\u6587\u5e93\u5feb\u901f\u7b5b\u9009\u4e0e\u8bbe\u8ba1\u7684\u5b9e\u64cd\u65b9\u6cd5\u3002<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[4],"tags":[],"class_list":["post-9660","post","type-post","status-publish","format-standard","hentry","category-note"],"_links":{"self":[{"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/posts\/9660","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/fanyiming.life\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=9660"}],"version-history":[{"count":2,"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/posts\/9660\/revisions"}],"predecessor-version":[{"id":13868,"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/posts\/9660\/revisions\/13868"}],"wp:attachment":[{"href":"https:\/\/fanyiming.life\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=9660"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/fanyiming.life\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=9660"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/fanyiming.life\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=9660"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}