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，是负责父母的起源特异性表达的基因。在11 p 15.5处的印记的破坏发展Beckwith-Wiedemann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.对日本BWS病例的分析显示，与北美和欧洲患者相比，H19-DMR超甲基化的频率显著降低，染色体异常的频率较高。这些结果表明，表观遗传和遗传改变的易感性在两组之间存在差异。使用含有人父体11号染色体的小鼠杂交细胞产生其中表达LIT 1的短转录物0.2 kb、1.1 kb和6.6 kb的截短细胞。qRT-PCR显示在这些细胞中印迹KvLQT 1基因的表达显著升高。结果表明，LIT 1基因的非编码转录本调控印迹基因的表达.为了鉴定一种新的基因组印记调节剂，我们试图产生特异性细胞，其能够被新霉素抗性检测到印记破坏。我们构建了一个敲入载体，其中IRES-neo盒插入印迹Kip 2基因的下游。将该载体转染到小鼠ES细胞中;然而，迄今为止没有获得重组克隆。因此，我们改变了载体的结构，我们现在正在筛选ES细胞。获得重组克隆后，我们将构建Kip 2-IRES-neo基因敲入小鼠并建立小鼠胚胎成纤维细胞。

项目成果

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

SNPアレイとH19-DMR CTCF6のメチル化分析により明らかにされたWT1異常型Wilms腫瘍の遺伝学的, 臨床的不均一性

SNP 阵列和 H19-DMR CTCF6 甲基化分析揭示 WT1 异常肾母细胞瘤的遗传和临床异质性

• 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
• 通讯作者: 金子 安比古

Wilms腫瘍におけるIGF2のloss of imprinting(LOI)とWT1構造異常

肾母细胞瘤中 IGF2 印记缺失 (LOI) 和 WT1 结构异常

• 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
• 通讯作者: 春田 雅之

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

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