Bidirectional approach for elucidation of genomic imprinting mechanism

阐明基因组印记机制的双向方法

• 批准号: 18590313
• 负责人: SOEJIMA Hidenobu
• 金额: $ 2.4万
• 依托单位: Saga University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590313/
• 关键词: genomic imprinting; KIP2; LIT1 imprinted domain; Beckwith-Wiedemann syndrome; non-coding RNA; non-coding RNA; non-codin RNA

Genomic imprinting is an epigenetic phenomenon that is responsible for parent-of-origin specific expression of genes. A disruption of imprinting at 11p 15.5 develops Beckwith-Wiedemann syndrome (BWS) and tumors. To clarify a molecular mechanism of genomic imprinting, we studied following issues.1. Analysis of Japanese cases of BWS revealed a significantly lower frequency of H19-DMR hypermethylation and a higher frequency of chromosome abnormality than in North American and European patients. These results suggest that susceptibility to epigenetic and genetic alterations differs between the two groups.2. Truncation cells in which short transcripts, 0.2 kb, 1.1 kb, and 6.6 kb, of LIT1were expressed were generated using a mouse hybrid cell containing human paternal chromosome 11. qRT-PCR revealed that expression of an imprinted KvLQT1 gene was drastically elevated in these cells. The result suggested that a non-coding transcript of LIT1 gene regulates expression of imprinted genes.3. To identify a novel regulator for genomic imprinting, we tried to generate specific cells, which are able to be detected an imprinting disruption by neomycin-resistance. We constructed a knock-in vector in which IRES-neo cassette were inserted into downstream of an imprinted Kip2 gene. The vector was transfected into mouse ES cells ; however, no recombinant clone was obtained to date. Thus, we changed a construction of the vector, and we are now screening of ES cells. After obtained recombinant clones, we will generate Kip2-IRES-neo knock-in mice and establish mouse embryonic fibroblast.

基因组印记是一种表观遗传现象，负责基因父母特异性表达。 11p 15.5的印迹的破坏会发展出贝克维斯·韦德曼综合征（BWS）和肿瘤。为了阐明基因组印记的分子机制，我们研究了以下问题。1。对日本BW病例的分析表明，与北美和欧洲患者相比，H19-DMR高甲基化的频率明显低于染色体异常的频率和较高的染色体异常。这些结果表明，对表观遗传和遗传改变的敏感性在两组之间有所不同。2。使用含有人类父亲染色体11的小鼠杂化细胞产生了表达的lit1-lit1-lit1-lit1-lit1-lit1-lit1-lit1-lit1以下截短细胞。QRT-PCR揭示了这些细胞中烙印的KVLQT1基因的表达显着升高。结果表明，LIT1基因的非编码转录器调节印迹基因的表达3。为了确定一种新型的基因组印记调节剂，我们试图产生特定的细胞，这些细胞能够被新霉素耐药性检测到烙印。我们构建了一个敲门载体，其中将IRES-NEO盒插入了一个印迹KIP2基因的下游。将载体转染到小鼠ES细胞中。但是，迄今为止尚无重组克隆。因此，我们改变了向量的构造，现在正在筛选ES细胞。获得重组克隆后，我们将生成KIP2-IRES-NEO敲入小鼠并建立小鼠胚胎成纤维细胞。

项目成果

Genetic and epigenetic alterations on the short arm of chromosome 11 are involved in a majority of sporadic Wilms' tumors

大多数散发性肾母细胞瘤均涉及 11 号染色体短臂上的遗传和表观遗传改变

• 发表时间: 2006
• 期刊: Brit J Cancer 95
• 作者: Yamasaki-Ishizaki;Y;Kayashima;T;Mapendano;CK;Soejima;H;Ohta;T;Masuzaki;H;Kinoshita;A;Urano;T;Yoshiura;KI;Matsumoto;N;Ishimaru;T;Mukai;T;Niikawa;N;Kishino;T;Sasaki K;Yamasaki・Ishizaki Y;副島 英伸;Yamada Y;Watanabe N;Satoh Y
• 通讯作者: Satoh Y

A case with partial trisomy of 11p15 transmitted from mother

母传11p15部分三体一例

• 发表时间: 2007
• 作者: Shimokawa;O.;Fu;R.;Soejima;H.;Sasaki;K.;Kondo;T.;Matsumoto;N.;Yoshiura;K.;Niikawa;N.;Harada;N
• 通讯作者: N

NSD1異常を伴うSotos症候群における11p15.5ゲノムインプリンティング領域の解析

伴NSD1异常的Sotos综合征11p15.5基因组印记区分析

• 发表时间: 2007
• 作者: Haruta;M.;Watanabe;N.;Nakadate;N.;Fukuzwa;M.;Soejima;H.;Kaneko;Y;春田 雅之;金子 安比古;近藤 雅人
• 通讯作者: 近藤 雅人

特集エピジェネティクスの最新テクノロジーヒストン修飾の個別およびゲノム網羅的解析法〜ChIP法とChIP on chip法

特色：表观遗传学的最新技术 组蛋白修饰的个体和全基因组分析方法 - ChIP 方法和 ChIP on Chip 方法

• 发表时间: 2007
• 期刊: バイオテクノロジージャーナル 7
• 作者: Morisaki T;Toyama K;Cheng J;Kawachi H;Shimizu F;Ikawa M;Okabe M;Morisaki H;Sasaki K;Watanabe N;Yamasaki-Ishizaki Y;副島 英伸
• 通讯作者: 副島 英伸

Bgckwith-Wiedemann症候群本邦例の包括的解析

日本Bgckwith-Wiedemann综合征病例综合分析

• 发表时间: 2006
• 作者: Haruta;M.;Watanabe;N.;Nakadate;N.;Fukuzwa;M.;Soejima;H.;Kaneko;Y;春田 雅之;金子 安比古;近藤 雅人;Soejima H;春田 雅之;副島 英伸
• 通讯作者: 副島 英伸