Development of novel retroviral vectors showing no gene silencing effect

开发无基因沉默效应的新型逆转录病毒载体

• 批准号: 22580104
• 负责人: MIYAKE Katsuhide
• 金额: $ 3.08万
• 依托单位: Ishikawa Prefectural University (2012)Nagoya University (2010-2011)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22580104/
• 关键词: SUMO; インテグラーゼ; レトロウイルス; PIASy; Pc2; MLV; PML; サイレンシング; HIV

Pulldown assay and coimmunoprecipitation of cell extracts in which the integrase or reverse transcriptase of Moloney murine leukemia virus was transiently expressed showed that both enzymes interacted with PML proteins. In infected cells, interaction between the integrase and PML was also observed. Transient expression of PIASy and SUMO proteins facilitated SUMOylation of the integrase but had no apparent effects on the interaction with PML. A FLAG-tagged integrase colocalized with PML protein possibly in the PML body. Knockdown of PML by small interfering RNA resulted in reduced viral cDNA levels and integration efficiency. This suggested that PML proteins activated reverse transcription. Furthermore, we discovered direct interaction between the integrase and polycomb protein, Pc2.

瞬时表达了莫洛尼鼠白血病病毒的整合酶或逆转录酶的细胞提取物的下拉分析和共免疫沉淀，表明这两种酶都与PML蛋白相互作用。在受感染的细胞中，还观察到积分酶和PML之间的相互作用。季节和相扑蛋白的瞬时表达促进了整合酶的Sumoylation，但对与PML的相互作用没有明显的影响。与PML蛋白一起在PML体内共定位的标记标记的集成酶。小型干扰RNA敲低PML导致病毒cDNA水平和整合效率降低。这表明PML蛋白激活了逆转录。此外，我们发现了整合酶和多肉液蛋白PC2之间的直接相互作用。

项目成果

トランスジェニックニワトリ作製のための新規DNA導入法の開発

开发一种新的DNA导入方法来生产转基因鸡

• 发表时间: 2012
• 作者: Ishida;S.;Sato;F.;Endo;T.;三宅克英 ・金岡英徳・西島謙一・飯島信司
• 通讯作者: 三宅克英 ・金岡英徳・西島謙一・飯島信司

Chiken oviduct-specific expression of transgene by a hybrid ovalbumin enhancer and the Tet expression system

通过混合卵清蛋白增强子和 Tet 表达系统对转基因进行鸡输卵管特异性表达

• 发表时间: 2012
• 期刊: J. Biosci. Bioeng
• 作者: D. Kodama;D. Nishimiya;K. Nishijima;Y. Okino;Y. Inayoshi;Y. Kojima;K. Ono;M. Motono;K. Miyake;Y. Kawabe;K. Kyogoku;T. Yamashita;M. Kamihira;and S. Iijima.
• 通讯作者: and S. Iijima.

レトロウイルスのゲノムインテグレーションメカニズムの解析

逆转录病毒基因组整合机制分析

• 发表时间: 2010
• 作者: 三宅克英;沖野雄気;稲吉勇仁;水谷昭文;西島謙一;飯島信司
• 通讯作者: 飯島信司

Transcription Factor YY1 Interacts with Retroviral Integrases and Facilitates Integration of Moloney Murine Leukemia Virus cDNA into the Host Chromosomes

• DOI: 10.1128/jvi.02681-09
• 发表时间: 2010-08-15
• 期刊: JOURNAL OF VIROLOGY
• 影响因子: 5.4
• 作者: Inayoshi, Yujin;Okino, Yuuki;Iijima, Shinji
• 通讯作者: Iijima, Shinji

Moloney murine leukemia virus integrase and reverse transcriptase interact with PML proteins

莫洛尼鼠白血病病毒整合酶和逆转录酶与 PML 蛋白相互作用

• 发表时间: 2012
• 期刊: J. Biochem
• 作者: Y. Okino;Y. Inayoshi;Y. Kojima;S.Kidani;H. Kaneoka;A. Honkawa;K. Nishijima;K. Miyake;and S. Iijima
• 通讯作者: and S. Iijima