{"id":18271,"date":"2021-04-27T10:02:04","date_gmt":"2021-04-27T01:02:04","guid":{"rendered":"http:\/\/mus.brc.riken.jp\/ja\/?page_id=18271"},"modified":"2021-10-15T11:32:57","modified_gmt":"2021-10-15T02:32:57","slug":"may_2021_mm","status":"publish","type":"page","link":"http:\/\/mus.brc.riken.jp\/ja\/mouse_of_month\/may_2021_mm","title":{"rendered":"\u52b9\u7387\u7684\u306a\u30b2\u30ce\u30e0\u7de8\u96c6\u30de\u30a6\u30b9\u4f5c\u88fd\u5b9f\u73fe\u306e\u305f\u3081 \u958b\u767a\u3055\u308c\u305fCas9 Tg\u30de\u30a6\u30b9"},"content":{"rendered":"\n\n\n
\"RIKEN<\/td>\nMay 2021<\/span>
\nMouse of the Month<\/span><\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\n

\u52b9\u7387\u7684\u306a\u30b2\u30ce\u30e0\u7de8\u96c6\u30de\u30a6\u30b9\u4f5c\u88fd\u5b9f\u73fe\u306e\u305f\u3081\u958b\u767a\u3055\u308c\u305f
Cas9 Tg\u30de\u30a6\u30b9<\/span><\/p>\n

\nB6;B6D2-Tg(CAG-hCas9)2Saku (RBRC10984)<\/a>
\n<\/h5>\n

\n\"sCAT<\/a><\/p>\n

Courtesy of Takayuki Sakurai, Ph.D.<\/p>\n

Production of genetically engineered mice using maCas9 zygotes.<\/p>\n

(A) Schematic illustration of the generation of maCas9 zygotes by IVF with sCAT Tg\/+ ♀ and +\/+ ♂. (B) Gel images of Cas9 genotyping and T7EI assay for each pup developed from maCas9 zygotes electroporated with only ramp1 gRNAs. Lanes 1-9 show PCR products amplified from the genetic DNA of 9 pups obtained. Pups 1, 4, 5, 7 and 9 were Tg\/+. Pups 2, 3, 6 and 8 were +\/+. All pups 1-9 were identified as indel mutants. M, lambda Hind III+100bp ladder markers. WC, wild control.<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\u73fe\u5728\u3001\u907a\u4f1d\u5b50\u6539\u5909\u30de\u30a6\u30b9\u306e\u4f5c\u88fd\u306b\u306f\u3001CRISPR\/Cas9\u30b7\u30b9\u30c6\u30e0\u3092\u7528\u3044\u305f\u30b2\u30ce\u30e0\u7de8\u96c6\u6cd5\u304c\u4e3b\u6d41\u306b\u306a\u308a\u3064\u3064\u3042\u308a\u307e\u3059\u3002\u3057\u304b\u3057\u3001\u5168\u3066\u306e\u907a\u4f1d\u5b50\u6539\u5909\u30de\u30a6\u30b9\u306e\u4f5c\u88fd\u304c\u3001\u3053\u306e\u65b9\u6cd5\u3067\u30ab\u30d0\u30fc\u3067\u304d\u308b\u308f\u3051\u3067\u306f\u3042\u308a\u307e\u305b\u3093\u3002\u5916\u6765DNA\u3092\u6a19\u7684\u90e8\u4f4d\u306b\u633f\u5165\u3059\u308b\u30ce\u30c3\u30af\u30a4\u30f3\u52b9\u7387\u304c\u4f4e\u304b\u3063\u305f\u308a\u3001\u540c\u6642\u306b\u8907\u6570\u306e\u907a\u4f1d\u5b50\u3092\u30bf\u30fc\u30b2\u30c3\u30c8\u306b\u7de8\u96c6\u3092\u52a0\u3048\u308b\u3053\u3068\u304c\u56f0\u96e3\u3067\u3042\u3063\u305f\u308a\u3068\u3001\u89e3\u6c7a\u3059\u3079\u304d\u554f\u984c\u70b9\u304c\u6b8b\u3063\u3066\u3044\u308b\u306e\u304c\u73fe\u72b6\u3067\u3059\u3002

\n\u4eca\u56de\u3054\u7d39\u4ecb\u3059\u308bsCAT (s<\/u>ystematically Ca<\/u>s9-expressing t<\/u>ransgenic)\u30de\u30a6\u30b9(RBRC10984)\u306f\u3001CAG \u30d7\u30ed\u30e2\u30fc\u30bf\u30fc\u5236\u5fa1\u4e0b\u3067humanized Cas9\u3092\u767a\u73fe\u3059\u308bTg\u30de\u30a6\u30b9\u3067\u3059[1]\u3002\u5bc4\u8a17\u8005\u306e\u685c\u4e95\u5148\u751f\u3089\u306f\u3001\u4e0a\u8a18\u306e\u554f\u984c\u70b9\u3092\u89e3\u6c7a\u3059\u308b\u305f\u3081\u306b\u3001\u52b9\u7387\u7684\u306b\u30b2\u30ce\u30e0\u7de8\u96c6\u30de\u30a6\u30b9\u3092\u4f5c\u88fd\u3059\u308b\u305f\u3081\u306e\u30c4\u30fc\u30eb\u3068\u3057\u3066sCAT\u30de\u30a6\u30b9\u3092\u958b\u767a\u3057\u307e\u3057\u305f\u3002\u305d\u3057\u3066\u3001sCAT\u96cc\u30de\u30a6\u30b9\u3092\u7528\u3044\u3066\u4f5c\u51fa\u3057\u305f\u4f53\u5916\u53d7\u7cbe\u80damaCas9 zygotes (ma<\/u>ternal Cas9<\/u>\u3092\u542b\u3080\u53d7\u7cbe\u5375)\u3092\u7528\u3044\u305f\u3001\u65b0\u898f\u30b2\u30ce\u30e0\u7de8\u96c6\u30de\u30a6\u30b9\u4f5c\u88fd\u65b9\u6cd5\u3092\u5831\u544a\u3057\u307e\u3057\u305f[2]\u3002\u307e\u305a\u3001\u672c\u65b9\u6cd5\u3067\u3001\u9ad8\u52b9\u7387\u306a\u30ce\u30c3\u30af\u30a4\u30f3\u30de\u30a6\u30b9\u4f5c\u88fd\u304c\u53ef\u80fd\u3067\u3042\u308b\u304b\u3092\u78ba\u8a8d\u3059\u308b\u305f\u3081\u3001\u5358\u4e00\u907a\u4f1d\u5b50\u3092\u30bf\u30fc\u30b2\u30c3\u30c8\u306b\u3001\u4efb\u610f\u306egRNA (g<\/u>uide RNA<\/u>)\u3068HDR (h<\/u>omology d<\/u>irect r<\/u>epair)\u306e\u30c6\u30f3\u30d7\u30ec\u30fc\u30c8\u3068\u306a\u308b\u30ce\u30c3\u30af\u30a4\u30f3\u7528\u306essODN (s<\/u>ingle-s<\/u>trand o<\/u>ligod<\/u>eoxyn<\/u>ucleotide)\u3092\u3001maCas9 zygotes\u306b\u30a8\u30ec\u30af\u30c8\u30ed\u30dd\u30ec\u30fc\u30b7\u30e7\u30f3\u6cd5\u306b\u3088\u3063\u3066\u3001\u5c0e\u5165\u3057\u307e\u3057\u305f\u3002\u305d\u306e\u7d50\u679c\u3001\u5f93\u6765\u306e\u5916\u6765Cas9 mRNA(\u307e\u305f\u306f\u30bf\u30f3\u30d1\u30af\u8cea)\u3092\u5171\u5c0e\u5165\u3057\u305f\u5834\u5408\u3068\u6bd4\u8f03\u3057\u3066\u3001\u30e2\u30b6\u30a4\u30af\u5909\u7570\u7387\u306f\u4f4e\u304f\u3001\u30ce\u30c3\u30af\u30a4\u30f3\u52b9\u7387\u306f\u9ad8\u304f\u3001\u30de\u30a6\u30b9\u51fa\u751f\u7387\u3082\u9ad8\u3044\u3068\u3044\u3046\u7d50\u679c\u3092\u5f97\u307e\u3057\u305f\u3002\u307e\u305f\u3001\u540c\u6642\u306b\u8907\u6570\u306e\u907a\u4f1d\u5b50\u3092\u30bf\u30fc\u30b2\u30c3\u30c8\u306b\u3059\u308b\u3053\u3068\u304c\u53ef\u80fd\u3067\u3042\u308b\u304b\u3092\u78ba\u8a8d\u3059\u308b\u305f\u3081\u306b\u300110\u907a\u4f1d\u5b50\u306b\u5bfe\u3059\u308bgRNA\u3092\u4e00\u5ea6\u306b\u30a8\u30ec\u30af\u30c8\u30ed\u30dd\u30ec\u30fc\u30b7\u30e7\u30f3\u6cd5\u306b\u3088\u3063\u3066maCas9 zygotes\u306b\u5c0e\u5165\u3057\u305f\u3068\u3053\u308d\u3001\u6700\u5927\u30679\u907a\u4f1d\u5b50\u304cindel (in<\/u>sertion\/del<\/u>etion)\u5909\u7570\u3092\u751f\u3058\u3066\u304a\u308a\u3001\u3053\u306e\u5834\u5408\u306b\u304a\u3044\u3066\u3082\u3001\u5f93\u6765\u306e\u65b9\u6cd5\u3068\u6bd4\u8f03\u3057\u3066\u3001indel\u5909\u7570\u7387\u306f\u9ad8\u304f\u3001\u30de\u30a6\u30b9\u51fa\u751f\u7387\u3082\u9ad8\u3044\u3068\u3044\u3046\u7d50\u679c\u3092\u5f97\u307e\u3057\u305f\u3002

\n\u3053\u308c\u3088\u308a\u3001maCas9 zygotes\u3092\u7528\u3044\u305f\u30b2\u30ce\u30e0\u7de8\u96c6\u6cd5\u306f\u3001\u52b9\u7387\u7684\u306a\u30ce\u30c3\u30af\u30a4\u30f3\u30de\u30a6\u30b9\u306e\u4f5c\u88fd\u3001\u304a\u3088\u3073\u3001\u591a\u91cd\u907a\u4f1d\u5b50\u30b2\u30ce\u30e0\u7de8\u96c6\u30de\u30a6\u30b9\u306e\u4f5c\u88fd\u306b\u6709\u7528\u3067\u3042\u308a\u3001\u4eca\u5f8c\u3001\u591a\u56e0\u5b50\u75be\u60a3\u30e2\u30c7\u30eb\u306e\u4f5c\u88fd\u7b49\u306b\u5fdc\u7528\u5229\u7528\u3055\u308c\u308b\u3053\u3068\u304c\u671f\u5f85\u3055\u308c\u307e\u3059\u3002\u307e\u305f\u3001sCAT\u30de\u30a6\u30b9\u306f\u3001\u5fc3\u81d3\u3001\u7cbe\u5de3\u3001\u7b4b\u306a\u3069\u4e3b\u8981\u306a\u81d3\u5668\u3067\u306eCas9 mRNA\u306e\u767a\u73fe\u304c\u78ba\u8a8d\u3055\u308c\u3066\u3044\u308b\u3053\u3068\u304b\u3089\u3001\u4ed6\u306ein vivo<\/i> \u304a\u3088\u3073in vitro<\/i>\u5b9f\u9a13\u306b\u304a\u3044\u3066\u3082\u3001\u6709\u7528\u3067\u3042\u308b\u3068\u3044\u3048\u307e\u3059\u3002<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n\n\n\n\n
Depositor<\/td>\n:<\/td>\n\u685c\u4e95 \u656c\u4e4b \u5148\u751f
\u4fe1\u5dde\u5927\u5b66\n<\/td>\n<\/tr>\n
Strain name<\/td>\n:<\/td>\nB6;B6D2-Tg(CAG-hCas9)2Saku<\/td>\n<\/tr>\n
RBRC No.<\/td>\n:<\/td>\nRBRC10984<\/td>\n<\/tr>\n
References<\/td>\n:<\/td>\n[1]<\/td>\nSakurai T, Kamiyoshi A, Kawate H, Mori C, Watanabe S, Tanaka M, Uetake R, Sato M, Shindo T.
\nA non-inheritable maternal Cas9-based multiple-gene editing system in mice.
\nSci Rep.<\/em> 2016 Jan 28;6:20011.
\n<\/a><\/td>\n<\/tr>\n
[2]<\/td>\nSakurai T, Kamiyoshi A, Kawate H, Watanabe S, Sato M, Shindo T.
\nProduction of genetically engineered mice with higher efficiency, lower mosaicism, and multiplexing capability using maternally expressed Cas9.
\nSci Rep.<\/em> 2020 Jan 23;10(1):1091.<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
May 2021
\nSaori Mizuno, Ph.D.
\nContact: Experimental Animal Division, RIKEN BioResource Research Center (animal.brc@riken.jp)<\/a>
\nAll materials contained on this site may not be reproduced, distributed, displayed, published or broadcast without the prior permission of the owner of that content.<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n","protected":false},"excerpt":{"rendered":"

May 2021 Mouse of the Month \u52b9\u7387\u7684\u306a\u30b2\u30ce\u30e0\u7de8\u96c6\u30de\u30a6\u30b9\u4f5c\u88fd\u5b9f\u73fe\u306e\u305f\u3081\u958b\u767a\u3055\u308c\u305fCas9 Tg\u30de\u30a6\u30b9 B6;B6D2-Tg(CAG-hCas9)2Saku (RBRC10984) Courtesy of Takayuki Sakurai, Ph.D. Production of genetically engineered mice using maCas9 zygotes […]<\/p>\n","protected":false},"author":19,"featured_media":0,"parent":198,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_seopress_titles_title":"","_seopress_titles_desc":"","_seopress_robots_index":"","_seopress_robots_follow":"","_seopress_robots_imageindex":"","_seopress_robots_snippet":"","_seopress_robots_primary_cat":"","_seopress_robots_breadcrumbs":"","_seopress_robots_freeze_modified_date":"","_seopress_robots_custom_modified_date":"","_seopress_robots_canonical":"","_seopress_social_fb_title":"","_seopress_social_fb_desc":"","_seopress_social_fb_img":"","_seopress_social_fb_img_attachment_id":0,"_seopress_social_fb_img_width":0,"_seopress_social_fb_img_height":0,"_seopress_social_twitter_title":"","_seopress_social_twitter_desc":"","_seopress_social_twitter_img":"","_seopress_social_twitter_img_attachment_id":0,"_seopress_social_twitter_img_width":0,"_seopress_social_twitter_img_height":0,"_seopress_redirections_value":"","_seopress_redirections_enabled":"","_seopress_redirections_enabled_regex":"","_seopress_redirections_logged_status":"","_seopress_redirections_param":"","_seopress_redirections_type":301,"_seopress_analysis_target_kw":"","footnotes":"","_wp_rev_ctl_limit":""},"class_list":["post-18271","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"http:\/\/mus.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/18271","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/mus.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"http:\/\/mus.brc.riken.jp\/ja\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"http:\/\/mus.brc.riken.jp\/ja\/wp-json\/wp\/v2\/users\/19"}],"replies":[{"embeddable":true,"href":"http:\/\/mus.brc.riken.jp\/ja\/wp-json\/wp\/v2\/comments?post=18271"}],"version-history":[{"count":7,"href":"http:\/\/mus.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/18271\/revisions"}],"predecessor-version":[{"id":18752,"href":"http:\/\/mus.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/18271\/revisions\/18752"}],"up":[{"embeddable":true,"href":"http:\/\/mus.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/198"}],"wp:attachment":[{"href":"http:\/\/mus.brc.riken.jp\/ja\/wp-json\/wp\/v2\/media?parent=18271"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}