Approach to allele specific and regulated gene silencing using the artificial miRNA expression system

使用人工 miRNA 表达系统实现等位基因特异性和调控基因沉默的方法

• 批准号: 21659084
• 负责人: FUKUMAKI Yasuyuki
• 金额: $ 2.07万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21659084/
• 关键词: マイクQRNA; マイクQR; 遺伝子変異; アレル特異的; マイクロRNA; 発現抑制; FAP; TTR; オフターゲット効果; ミスマッチ

The allele specific and regulated gene silencing is needed in various aspects of biological and medical fields. To establish this system, artificial miRNA driven by the poly II promoter was designed for discrimination between normal and mutated alleles in silencing effect. Although the allele specific silencing was obtained using artificial miRNA, there was no increased silencing effect expected to be provided by co-expression of another miRNA targeting the region shared by two genotypes. Cre dependent silencing of Smad2 and Smad3 as a model was obtained in ES cells using the construct in which expression of two miRNA species is blocked by the Neo^r gene flanked by two loxP sites.

在生物和医学领域的各个方面都需要等位基因特异的、受调控的基因沉默。为了建立这一体系，设计了由PolyII启动子驱动的人工miRNA来区分沉默效应中正常和突变的等位基因。虽然利用人工miRNA获得了等位基因特异性沉默，但针对两个基因型共有的区域的另一种miRNA的共表达并没有增加沉默效果。在ES细胞中，利用两个miRNA物种的表达被Neo^r基因两侧的两个loxP位点阻断的结构，获得了依赖Cre的Smad2和Smad3的沉默模型。

项目成果

Multiple system degeneration with basophilic inclusions in Japanese ALS patients with FUS mutation

• DOI: 10.1007/s00401-009-0621-1
• 发表时间: 2010-03-01
• 期刊: ACTA NEUROPATHOLOGICA
• 影响因子: 12.7
• 作者: Tateishi, Takahisa;Hokonohara, Toshihiro;Kira, Jun-ichi
• 通讯作者: Kira, Jun-ichi

Partial SPAST and DPY30 deletions in a Japanese spastic paraplegia type 4 family

日本痉挛性截瘫 4 型家族中 SPAST 和 DPY30 部分缺失

• 发表时间: 2011
• 期刊: Neurogenetics
• 影响因子: 2.2
• 作者: Miura;S.;Shibata;H.;Kida;H.;Noda;K.;Toyama;T.;Iwasaki;N.;Iwaki;A.;Ayabe;M.;Aizawa;H.;Taniwaki;T. and Fukumaki;Y.
• 通讯作者: Y.

Linkageassisted exome sequencing to identify the responsible variant for a novel type of hereditary motor and sensory neuropathy with proximal dominancy in the lower extremities found in a single Japanese family

连锁辅助外显子组测序可识别日本一个家庭中发现的一种新型遗传性运动和感觉神经病的致病变异，该病以下肢近端为主

• 发表时间: 2012
• 作者: Shibata;H.;Miura;S.;Kida;H.;Neda;K.;Kaku;Y.;Iwaki;A.;Ayabe;M.;Taniwaki;T. and Fukumaki;Y.
• 通讯作者: Y.

Linkage-assisted exome sequencing to identify the responsible variant for a novel type of hereditary motor and sensory neuropathy with proximal dominancy in the lower extremities found in a single Japanese family

连锁辅助外显子组测序可鉴定日本一个家庭中发现的一种新型遗传性运动和感觉神经病的致病变异，该病以下肢近端为主

• 发表时间: 2012
• 作者: Shibata;H.;et al
• 通讯作者: et al

A novel type of hereditary motor and sensory neuropathy with proximal dominancy in the lower extremities found in a Japanese descent

日本血统中发现的一种新型遗传性运动和感觉神经病，下肢近端优势

• 发表时间: 2010
• 作者: Miura;S.;Shibata;H.;Neda;K.;Y. Kaku;Y. Kida;H.;Iwaki;A.;Ayabe;M.;Aizawa;H.;Taniwaki;T. and Fukumaki;Y.
• 通讯作者: Y.