{"id":1249,"date":"2018-09-30T00:00:00","date_gmt":"2018-09-29T15:00:00","guid":{"rendered":"https:\/\/www.thermofisher.com\/blog\/learning-at-the-bench\/user_voice_36_4_lipofectamine\/"},"modified":"2023-12-28T09:21:36","modified_gmt":"2023-12-28T00:21:36","slug":"user_voice_36_4_lipofectamine","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/learning-at-the-bench\/user_voice_36_4_lipofectamine\/","title":{"rendered":"\u3055\u3089\u306b\u9032\u5316\u3057\u3001\u6c4e\u7528\u5316\u3059\u308bCRISPR-Cas9\u30b7\u30b9\u30c6\u30e0\u3078\uff08\u5e83\u5cf6\u5927\u5b66\u5927\u5b66 \u5206\u5b50\u907a\u4f1d\u5b66\u7814\u7a76\u5ba4 \u5c71\u672c\u5353\u6c0f\u3001\u4f50\u4e45\u9593\u54f2\u53f2\u6c0f\uff09-\u300cNEXT\u300d2016\u5e747\u6708\u53f7\u63b2\u8f09"},"content":{"rendered":"

RNP\u5c0e\u5165\u306bLipofectamine CRISPRMAX\u3092\u6d3b\u7528
\n<\/span><\/h2>\n

\u5c71\u672c\u5353\u6c0f\uff08\u5e83\u5cf6\u5927\u5b66\u5927\u5b66\u9662\u7406\u5b66\u7814\u7a76\u79d1\u6570\u7406\u5206\u5b50\u751f\u547d\u7406\u5b66\u5c02\u653b\u5206\u5b50\u907a\u4f1d\u5b66\u7814\u7a76\u5ba4\u30fb\u6559\u6388\uff09<\/strong>
\n\u4f50\u4e45\u9593\u54f2\u53f2\u6c0f\uff08\u540c\u30fb\u7279\u4efb\u8b1b\u5e2b\uff09
\n<\/strong><\/p>\n

\"\u5c71\u672c\u5353\u6c0f\uff08\u5e83\u5cf6\u5927\u5b66\u5927\u5b66\u9662\u7406\u5b66\u7814\u7a76\u79d1\u6570\u7406\u5206\u5b50\u751f\u547d\u7406\u5b66\u5c02\u653b\u5206\u5b50\u907a\u4f1d\u5b66\u7814\u7a76\u5ba4\u30fb\u6559\u6388\uff09\"<\/a>\u30b2\u30ce\u30e0\u7de8\u96c6\u306e\u9ece\u660e\u671f\u304b\u3089\u6280\u8853\u958b\u767a\u3092\u727d\u5f15\u3057\u3001\u5b66\u4f1a\u3092\u7acb\u3061\u4e0a\u3052\u3001\u56fd\u5185\u3078\u306e\u6d78\u900f\u306b\u8ca2\u732e\u3059\u308b\u5e83\u5cf6\u5927\u5b66\u306e\u5c71\u672c\u5353\u6c0f\u3068\u4f50\u4e45\u9593\u54f2\u53f2\u6c0f\u3002\u4f7f\u3044\u3084\u3059\u304f\u3001\u3055\u3089\u306b\u5fdc\u7528\u7bc4\u56f2\u3092\u5e83\u3052\u308bCRISPR-Cas9\u30b7\u30b9\u30c6\u30e0\u306e\u6280\u8853\u958b\u767a\u3084\u4eca\u5f8c\u306e\u767a\u5c55\u306b\u3064\u3044\u3066\u4f3a\u3044\u307e\u3057\u305f\u3002<\/p>\n

\u7d30\u80de\u5c0e\u5165\u6cd5\u3067\u7de8\u96c6\u52b9\u7387\u3092\u4e0a\u3052\u308b<\/span><\/h2>\n

\u300cZFN\u3001TALEN\u306b\u7d9a\u3044\u3066\u958b\u767a\u3055\u308c\u305fCRISPR-Cas9\u6280\u8853\u306b\u3088\u308a\u3001\u30b2\u30ce\u30e0\u7de8\u96c6\u306f\u3088\u308a\u8eab\u8fd1\u306b\u306a\u308a\u307e\u3057\u305f\u3002\u7279\u306bCas9\u30cc\u30af\u30ec\u30a2\u30fc\u30bc\u30bf\u30f3\u30d1\u30af\u8cea\u3092\u5229\u7528\u3059\u308b\u3053\u3068\u3067\u3053\u306e\u30b7\u30b9\u30c6\u30e0\u306e\u5fdc\u7528\u6027\u306f\u3055\u3089\u306b\u5e83\u304c\u308a\u305d\u3046\u300d\u3068\u5c71\u672c\u6c0f\u3002gRNA\uff08\u30ac\u30a4\u30c9RNA\uff09\u3084Cas9\u914d\u5217\u3092\u30d7\u30e9\u30b9\u30df\u30c9\u3084mRNA\u3067\u5171\u5c0e\u5165\u3059\u308b\u5f93\u6765\u6cd5\u306b\u6bd4\u3079\u3001Cas9\u30bf\u30f3\u30d1\u30af\u8cea\u3068gRNA\u3092RNP\uff08Ribonucleo-protein\uff09\u3068\u3057\u3066\u5c0e\u5165\u3059\u308b\u3068\u3001DNA\u5207\u65ad\u307e\u3067\u306e\u6642\u9593\u304c\u77ed\u304f\u3001\u904e\u5270\u767a\u73fe\u306b\u3088\u308b\u30aa\u30d5\u30bf\u30fc\u30b2\u30c3\u30c8\u5207\u65ad\u306e\u5371\u967a\u6027\u304c\u4f4e\u3044\u4e0a\u3001\u591a\u304f\u306e\u7d30\u80de\u3067\u7de8\u96c6\u52b9\u7387\u304c\u5411\u4e0a\u3057\u307e\u3059\u3002\u300c\u57f9\u990a\u7d30\u80de\u3078\u306eRNP\u5c0e\u5165\u306b\u306f\u300eLipofectamine CRISPRMAX\u300f\u3092\u4f7f\u3063\u3066\u3044\u307e\u3059\u3002\u30a8\u30ec\u30af\u30c8\u30ed\u30dd\u30ec\u30fc\u30b7\u30e7\u30f3\u6cd5\u306e\u3088\u3046\u306b\u5c02\u7528\u6a5f\u5668\u3082\u5fc5\u8981\u306a\u304f\u30e9\u30f3\u30cb\u30f3\u30b0\u30b3\u30b9\u30c8\u304c\u6291\u3048\u3089\u308c\u307e\u3059\u3002\u307e\u305f\u4ed6\u306e\u5c0e\u5165\u6cd5\u3067\u306f\u52b9\u7387\u304c\u4f4e\u3044HCT116\u682a\u3084HepG2\u682a\u3067\u306e\u52b9\u7387\u3082\u4e0a\u304c\u308a\u307e\u3057\u305f\u300d\u3068\u4f50\u4e45\u9593\u6c0f\u3082\u30b3\u30e1\u30f3\u30c8\u3057\u307e\u3059\u3002\u300c\u3057\u304b\u3082Cas9\u30bf\u30f3\u30d1\u30af\u8cea\u3092\u4f7f\u3046\u7cfb\u3067\u306f\u3001\u4e8b\u524d\u306egRNA\u306e\u8a55\u4fa1\u3082\u7c21\u5358\u3067\u3059\u3002in vitro<\/i>\u3067\u3001\u6a19\u7684\u914d\u5217\u3092\u542b\u3080DNA\u65ad\u7247\u306bgRNA\u3068Cas9\u30bf\u30f3\u30d1\u30af\u8cea\u3092\u53cd\u5fdc\u3055\u305b\u3066\u3001\u5207\u65ad\u6d3b\u6027\u306e\u9ad8\u3044gRNA\u3092\u9078\u5225\u3067\u304d\u308b\u306e\u3067\u300d\u3068\u5c71\u672c\u6c0f\u3002\u300c\u3055\u3089\u306b\u7de8\u96c6\u5f8c\u306e\u8a55\u4fa1\u306b\u306f\u3001\u30d7\u30e9\u30b9\u30df\u30c9\u306e\u5834\u5408\u306fInvitrogen\u2122 GeneArt\u2122 Genomic Cleavage Detection Kit\u3092\u4f7f\u3044\u307e\u3059\u304c\u3001Cas9\u30bf\u30f3\u30d1\u30af\u8cea\u3092\u4f7f\u3046\u7cfb\u3067\u306f\u3001\u7de8\u96c6\u5f8c\u306e\u7d30\u80de\u96c6\u56e3\u304b\u3089\u30b2\u30ce\u30e0DNA\u3092\u62bd\u51fa\u3057\u3001\u540c\u3058\u304f\u6a19\u7684\u90e8\u4f4d\u3092\u542b\u3080\u9818\u57df\u3092PCR\u5897\u5e45\u3055\u305b\u3066in vitro<\/i>\u3067\u5207\u65ad\u53cd\u5fdc\u3092\u884c\u3044\u307e\u3059\u3002\u914d\u5217\u304c\u5909\u5316\u3057\u3066\u3044\u308c\u3070\u5207\u65ad\u3055\u308c\u305a\u3001\u7de8\u96c6\u304c\u8d77\u304d\u305f\u7d30\u80de\u306e\u5272\u5408\u304c\u5206\u304b\u308a\u307e\u3059\u3002\u5b9f\u969b\u306e\u52b9\u7387\u306f\u3001\u30b2\u30ce\u30e0\u306e\u72b6\u614b\u3084\u907a\u4f1d\u5b50\u5ea7\u306b\u3088\u3063\u3066\u7570\u306a\u308b\u306e\u3067\u6709\u7528\u3067\u3059\u3002\u3053\u308c\u304b\u3089\u306fCas9\u30bf\u30f3\u30d1\u30af\u8cea\u3092\u5236\u9650\u9175\u7d20\u306e\u3088\u3046\u306b\u4f7f\u3048\u305d\u3046\u300d\u3068\u7d9a\u3051\u307e\u3059\u3002<\/p>\n

\u52b9\u7387\u3088\u3044\u30ce\u30c3\u30af\u30a4\u30f3\u6cd5\u306e\u666e\u53ca\u3092\u76ee\u6307\u3057\u3066<\/span><\/h2>\n

\u300c\u30b2\u30ce\u30e0\u7de8\u96c6\u3092\u5229\u7528\u3057\u305f\u30ce\u30c3\u30af\u30a2\u30a6\u30c8\u6280\u8853\u306f\u3001\u3059\u3067\u306b\u6c4e\u7528\u7684\u306a\u624b\u6cd5\u3068\u3057\u3066\u5e83\u304c\u3063\u3066\u3044\u307e\u3059\u3002\u6b21\u306f\u30ce\u30c3\u30af\u30a4\u30f3\u52b9\u7387\u3092\u4e0a\u3052\u3066\u8ab0\u3082\u304c\u4f7f\u3048\u308b\u6280\u8853\u306b\u3057\u305f\u3044\u3067\u3059\u306d\u300d\u3068\u5c71\u672c\u6c0f\u3002\u5f93\u6765\u6cd5\u3067\u306f500~1000\u5869\u57fa\u5bfe\u3068\u3044\u3046\u9577\u3044\u76f8\u540c\u9818\u57df\u3092\u6301\u3064\u30c9\u30ca\u30fc\u30d9\u30af\u30bf\u30fc\u3092\u4f5c\u88fd\u3057\u3066\u76f8\u540c\u7d44\u63db\u3048\u3092\u8d77\u3053\u3059\u65b9\u6cd5\u3068\u3001\u30b2\u30ce\u30e0\u7de8\u96c6\u306e\u5207\u65ad\u7b87\u6240\u306b\u3059\u3063\u307d\u308a\u3068DNA\u914d\u5217\u3092\u7d44\u307f\u8fbc\u3080\u65b9\u6cd5\u304c\u3042\u308a\u307e\u3057\u305f\u3002\u3057\u304b\u3057\u3001\u524d\u8005\u306f\u4fee\u5fa9\u6d3b\u6027\u306e\u5f31\u3044\u7d30\u80de\u3067\u306f\u52b9\u7387\u304c\u4f4e\u304f\u3001\u5f8c\u8005\u306f\u7d44\u307f\u8fbc\u3080\u5411\u304d\u304c\u5236\u5fa1\u3067\u304d\u307e\u305b\u3093\u3002\u305d\u3053\u3067\u958b\u767a\u3057\u305f\u306e\u304c\u300cPITCh\uff08Precise Integration into Target Chromosome\uff09\u6cd5\u300d\u3067\u3059\u3002\u300cCas9\u3067\u3001\u30b2\u30ce\u30e0DNA\u3068\u30c9\u30ca\u30fc\u30d9\u30af\u30bf\u30fc\u306e\u633f\u5165\u907a\u4f1d\u5b50\u306e\u524d\u5f8c\u306e3\u30ab\u6240\u3092\u5207\u65ad\u3057\u300120\u5869\u57fa\u5bfe\u307b\u3069\u306e\u77ed\u3044\u76f8\u540c\u914d\u5217\u3092\u624b\u304c\u304b\u308a\u306b\u3001\u30de\u30a4\u30af\u30ed\u30db\u30e2\u30ed\u30b8\u30fc\u5a92\u4ecb\u672b\u7aef\u7d50\u5408\u3068\u3044\u3046\u4fee\u5fa9\u6a5f\u69cb\u3067\u5916\u6765\u907a\u4f1d\u5b50\u3092\u30ce\u30c3\u30af\u30a4\u30f3\u3057\u307e\u3059\u3002\u76f8\u540c\u7d44\u63db\u3048\u3088\u308a\u6b63\u78ba\u6027\u306f\u3044\u304f\u3089\u304b\u843d\u3061\u307e\u3059\u304c\u3001\u624b\u8efd\u3067\u52b9\u7387\u304c\u5411\u4e0a\u3057\u307e\u3059\u3002\u3053\u308c\u307e\u3067\u52b9\u7387\u304c\u4f4e\u304b\u3063\u305f\u30ab\u30a8\u30eb\u3084\u30bc\u30d6\u30e9\u30d5\u30a3\u30c3\u30b7\u30e5\u3067\u3082\u30ce\u30c3\u30af\u30a4\u30f3\u500b\u4f53\u304c\u3068\u308c\u307e\u3059\u3088\u300d\u3068\u3001\u4f50\u4e45\u9593\u6c0f\u306f\u65b0\u3057\u3044\u6280\u8853\u958b\u767a\u306e\u30e1\u30ea\u30c3\u30c8\u3092\u8a71\u3057\u307e\u3059\u3002<\/p>\n

\u30b2\u30ce\u30e0\u7de8\u96c6\u7814\u7a76\u306e\u3053\u308c\u304b\u3089<\/span><\/h2>\n

\u5c71\u672c\u6c0f\u3068\u4f50\u4e45\u9593\u6c0f\u304c\u3001\u4eca\u5f8c\u6ce8\u76ee\u3059\u308b\u306e\u306f\u533b\u7642\u5206\u91ce\u3002\u300c\u75be\u60a3\u306f\u9650\u5b9a\u3055\u308c\u307e\u3059\u304c\u3001\u75be\u60a3\u90e8\u4f4d\u306e\u7d30\u80de\u3092\u4f53\u5916\u3067\u30b2\u30ce\u30e0\u7de8\u96c6\u3057\u3001\u7d30\u80de\u3092\u623b\u3059\u6cbb\u7642\u6cd5\u306a\u3069\u671f\u5f85\u3067\u304d\u305d\u3046\u3067\u3059\u300d\u3068\u8a9e\u308a\u307e\u3059\u3002\u307e\u305f\u5207\u65ad\u6d3b\u6027\u3092\u55aa\u5931\u3055\u305b\u305f\u30c7\u30c3\u30c9\u578bCas9\u3092\u3001\u8ee2\u5199\u56e0\u5b50\u3084\u86cd\u5149\u30bf\u30f3\u30d1\u30af\u8cea\u306a\u3069\u597d\u307f\u306e\u30bf\u30f3\u30d1\u30af\u8cea\u306b\u878d\u5408\u3055\u305b\u308b\u3053\u3068\u3067\u3001\u30b2\u30ce\u30e0\u4e0a\u306e\u72d9\u3063\u305f\u4f4d\u7f6e\u306b\u6d3b\u6027\u5206\u5b50\u3084\u30e2\u30cb\u30bf\u30fc\u5206\u5b50\u3092\u904b\u3076\u300c\u8239\u982d\u300d\u306e\u5f79\u5272\u306b\u3082\u671f\u5f85\u3057\u3066\u3044\u307e\u3059\u3002\u300c\u30af\u30ed\u30de\u30c1\u30f3\u30ea\u30e2\u30c7\u30ea\u30f3\u30b0\u56e0\u5b50\u3092\u878d\u5408\u3055\u305b\u3001DNA\u3092\u50b7\u3064\u3051\u305a\u306b\u30a8\u30d4\u30b2\u30ce\u30e0\u74b0\u5883\u3060\u3051\u3092\u5909\u3048\u3001\u907a\u4f1d\u5b50\u5ea7\u7279\u7570\u7684\u306b\u8ee2\u5199\u3092\u5236\u5fa1\u3059\u308b\u300e\u30a8\u30d4\u30b2\u30ce\u30e0\u7de8\u96c6\u300f\u304c\u9032\u3093\u3067\u3044\u307e\u3059\u3002\u304c\u3093\u306e\u5c02\u9580\u5bb6\u3068\u5354\u529b\u3057\u3001\u30a8\u30d4\u30b2\u30ce\u30e0\u306e\u7570\u5e38\u304c\u539f\u56e0\u3067\u8d77\u3053\u308b\u304c\u3093\u306e\u6cbb\u7642\u6cd5\u958b\u767a\u306b\u3082\u8ca2\u732e\u3057\u305f\u3044\u300d\u3068\u8a9e\u308a\u307e\u3059\u3002\u69d8\u3005\u306a\u5206\u91ce\u306e\u5c02\u9580\u5bb6\u3068\u306e\u30cd\u30c3\u30c8\u30ef\u30fc\u30af\u3092\u4ecb\u3057\u3066\u3001\u30b2\u30ce\u30e0\u7de8\u96c6\u6280\u8853\u306f\u591a\u69d8\u5316\u3057\u3064\u3064\u6d78\u900f\u3057\u3066\u3044\u304f\u3088\u3046\u3067\u3059\u3002<\/p>\n

\u30b2\u30ce\u30e0\u7de8\u96c6\u52b9\u7387\u3092\u6700\u5927\u5316! \u30bf\u30f3\u30d1\u30af\u8cea\u30d5\u30a9\u30fc\u30de\u30c3\u30c8\u3067\u306e\u5c0e\u5165\u306b\u6700\u9069
\nLipofectamine CRISPRMAX Cas9 Transfection Reagent<\/span><\/h2>\n

\"Lipofectamine<\/a>Invitrogen\u2122 Lipofectamine\u2122 CRISPRMAX\u2122 Cas9 Transfection Reagent\u306f\u3001Cas9 Nuclease\u30bf\u30f3\u30d1\u30af\u8cea\u3068RNA\u306e\u8907\u5408\u4f53\u3092\u9ad8\u3044\u52b9\u7387\u3067\u7d30\u80de\u306b\u5c0e\u5165\u3059\u308b\u8a66\u85ac\u3067\u3059\u3002\u8102\u8cea\u30d9\u30fc\u30b9\u306e\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u8a66\u85ac\u3068\u3057\u3066\u5b9a\u8a55\u3042\u308bLipofectamine\u30b7\u30ea\u30fc\u30ba\u3068\u3057\u3066\u958b\u767a\u3055\u308c\u307e\u3057\u305f\u3002<\/p>\n

Lipofectamine CRISPRMAX Cas9 Transfection Reagent\u306e\u8a73\u7d30\u60c5\u5831\u306f\u3053\u3061\u3089\u304b\u3089<\/a><\/p>\n

\u30e9\u30a4\u30d5\u30b5\u30a4\u30a8\u30f3\u30b9\u60c5\u5831\u8a8c\u300cNEXT\u300d<\/span><\/h2>\n

\"NEXT2016\u5e747\u6708\u53f7\"<\/a>\u5f53\u8a18\u4e8b\u306f\u30b5\u30fc\u30e2\u30d5\u30a3\u30c3\u30b7\u30e3\u30fc\u30b5\u30a4\u30a8\u30f3\u30c6\u30a3\u30d5\u30a3\u30c3\u30af\u304c\u767a\u520a\u3059\u308b\u30e9\u30a4\u30d5\u30b5\u30a4\u30a8\u30f3\u30b9\u60c5\u5831\u8a8c\u300cNEXT\u300d2016\u5e747\u6708\u53f7\u304b\u3089\u306e\u629c\u7c8b\u3067\u3059\u3002
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 <\/p>\n

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