J1 [129S4/SvJae] <a href='https://brc.riken.jp/mus/pcr03730'>Genotyping protocol -PCR-</a> Massachusetts General Hospital・岡野正樹先生、En Li（1999）。J1 ES細胞を用いて作出。 C (3-6 months) 岡野　正樹 Necessary documents for ordering:<ol><li>Order form (<A HREF="https://mus.brc.riken.jp/ja/wp-content/uploads/form/form_4.docx">Japanese</A> / <A HREF="https://mus.brc.riken.jp/en/wp-content/uploads/form/form_b.docx">English</A>)</li><li>Please use the MATERIAL TRANSFER AGREEMENT for Distribution <A HREF="https://mus.brc.riken.jp/en/wp-content/uploads/form/form_c.docx"> for non-profit</A> or <A HREF="https://mus.brc.riken.jp/en/wp-content/uploads/form/form_c_P.docx"> for profit</A> (English only).</li><li>Acceptance of responsibility for living modified organism (<A HREF="https://mus.brc.riken.jp/ja/wp-content/uploads/form/form_7.docx">Japanese</A> / <A HREF="https://mus.brc.riken.jp/en/wp-content/uploads/form/form_g.docx">English</A>)</li></ol> B6;129S4-Dnmt3a<tm1Enl> B6;129S4-Dnmt3a<tm1Enl> Masaki OKANO 条件を付加する。<br>A. RECIPIENT must send a copy of the executed MTA between RECIPIENT and RIKEN BRC to: Massachusetts General Hospital c/o Partners Innovation, Attn: TAG/MGH MTA 2020A005592, 399 Revolution Drive/Ste 950E, Somerville,MA 02245, USA (phsmta@partners.org). B. Any publication or public disclosure of research results obtained by the use of the BIOLOGICAL RESOURCE shall cite Massachusetts General Hospital in Boston, MA as the source of the material. However, neither the name, trademark, service mark, logo nor other identifying characteristic ("Name") of MGH or any of its affiliates, or any of its or their respective directors, trustees, officers, appointees, employees, staff, representatives or agents, in any advertising, promotional or sales literature, publicity or in any document employed to obtain funds or financing without the prior written approval of the MGH Department of Public Affairs. C. In publishing research results obtained by the use of the BIOLOGICAL RESOURCE, a citation of the literature Cell 1999 99:247-57 as designated by the DEPOSITOR is required. D. The use of the BIOLOGICAL RESOURCE is restricted to academic researchers in non-profit organizations for their internal research and educational purposes. E. The BIOLOGICAL RESOURCE shall not be used for commercial purposes. Any request for the BIOLOGICAL RESOURCE by a for-profit entity shall be referred to Massachusetts General Hospital through the Research and Licensing Office. F. Recipients shall assume all liability for their use, storage, handling and disposal of the BIOLOGICAL RESOURCE. The General Hospital Corporation d/b/a Massachusetts General Hospital will not be liable to the Recipients for any loss, claims, matters, damages, costs or liabilities relating to any third party actions, proceedings, investigations, or matters arising from any use, storage, handling, or disposal of the BIOLOGICAL RESOURCE by Recipient. <a href="https://mus.brc.riken.jp/ja/wp-content/uploads/pdf/blc/03730_BLC.pdf">Breeding characters</a> Heterozygote x Wild-type [C57BL/6JJcl] Dnmt3a遺伝子のノックアウトマウス。エクソン18内のApal siteからエクソン19内のNrul siteまでのゲノム領域 (DNAメチル化酵素活性中心PCおよびENVモチーフを含む) を、プロモーターを含まないIRES-βgeoカセットで置換。哺乳類は、互いに高いアミノ酸配列類似性のある二つのDe novo型DNAメチル化酵素Dnmt3a、Dnmt3bを持つ。両者は異なるアイソフォーム、発現パターンを示し、互いに重複する機能とそれぞれに特異的な機能を持つ。ノックアウトマウスでは、内在性Dnmt3aプロモーターからDnmt3a/IRES-βgeo融合転写物が発現、IRES配列によりlacZ/neo融合蛋白質が翻訳発現。Dnmt3a蛋白質産物はウェスタンブロットで検出されず、不安定な部分蛋白質は速やかに分解されると考えられる。DNAメチル化酵素活性中心領域を欠失することと考えあわせ、Dnmt3a完全欠失 (null) 変異と考えられる。ホモマウスは正常にうまれてくるものの、小型で、約4週齢で死亡する。Dnmt3a floxedマウス (RBRC03731) 、Dnmt3bノックアウトマウス (RBRC03732) 、Dnmt3b floxedマウス (RBRC03733) 。 Heterozygote x Wild-type [C57BL/6JJcl] true Dnmt3a knockout mice. A 0.3 kb region containing exons of Dnmt3a encoding PC and ENV motif was replaced with an IRES-beta-geo cassette. Homozygous mutant mice appear normal at birth, but most mutant mice become runted and die at about 4 week of age. Dnmt3a floxed mice (RBRC03731), Dnmt3b knockout mice (RBRC03732), Dnmt3b floxed mice (RBRC03733). Developed by Masaki Okano and En Li, Massachusetts General Hospital in 1999. J1 ES cells were used to generate the mutant mice. The mutant mice were crossed to C57BL/6. mouse En-2 splice acceptor, Encephalomyocarditis virus (EMCV) internal ribosomal entry site (ires), E. coli LacZ, E. coli neomycin registance gene, SV40 poly A signal sequence, mouse Dnmt3a genomic DNA C（3〜6か月） Dnmt3a<tm1Enl>; Dnmt3a null KO Dnmt3a<tm1Enl>; Dnmt3a null KO A. RECIPIENT must send a copy of the executed MTA between RECIPIENT and RIKEN BRC to: Massachusetts General Hospital c/o Partners Innovation, Attn: TAG/MGH MTA 2020A005592, 399 Revolution Drive/Ste 950E, Somerville,MA 02245, USA (phsmta@partners.org). B. Any publication or public disclosure of research results obtained by the use of the BIOLOGICAL RESOURCE shall cite Massachusetts General Hospital in Boston, MA as the source of the material. However, neither the name, trademark, service mark, logo nor other identifying characteristic ("Name") of MGH or any of its affiliates, or any of its or their respective directors, trustees, officers, appointees, employees, staff, representatives or agents, in any advertising, promotional or sales literature, publicity or in any document employed to obtain funds or financing without the prior written approval of the MGH Department of Public Affairs. C. In publishing research results obtained by the use of the BIOLOGICAL RESOURCE, a citation of the literature Cell 1999 99:247-57 as designated by the DEPOSITOR is required. D. The use of the BIOLOGICAL RESOURCE is restricted to academic researchers in non-profit organizations for their internal research and educational purposes. E. The BIOLOGICAL RESOURCE shall not be used for commercial purposes. Any request for the BIOLOGICAL RESOURCE by a for-profit entity shall be referred to Massachusetts General Hospital through the Research and Licensing Office. F. Recipients shall assume all liability for their use, storage, handling and disposal of the BIOLOGICAL RESOURCE. The General Hospital Corporation d/b/a Massachusetts General Hospital will not be liable to the Recipients for any loss, claims, matters, damages, costs or liabilities relating to any third party actions, proceedings, investigations, or matters arising from any use, storage, handling, or disposal of the BIOLOGICAL RESOURCE by Recipient. RBRC03730