Development of gene functional analysis system using human artificial chromosome in mouse

利用人类人工染色体的小鼠基因功能分析系统的开发

• 批准号: 23700519
• 负责人: OKAMURA Yoshiaki
• 金额: $ 2.75万
• 依托单位: 公益財団法人かずさDNA 研究所 (2012)Kazusa DNA Research Institute (2011)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23700519/
• 关键词: 動物実験技術; ヒト人工染色体; トランスジェニックマウス; 遺伝子機能解析手法開発

Human Artificial Chromosome (HAC) is a chromosomal vector that has repetitive DNA sequence derived from chentromere region, which can be segregated independently with host chromosome and stably maintained within the cells. Recently, we demonstrated that HAC introduced tetO sequences (tetO-HAC) can be eliminated from cells by inducing excess heterochromatin using transcriptional repressor, tTS. To develop the gene functional analysis system using tetO-HAC in mouse, we generated tetO-HAC that can be integrated gene expression cassettes. Then, GFP or tTS-EYFP expression cassettes was integrated into the tetO-HAC. In the case of the tetO-HAC that has tTS-EYFP cassette, tetO-HAC was eliminated from cells in the absence of Dox, which induce heterochromatin into the tetO-HAC. These data provide an important basis for development of the gene functional analysis system using tetO-HAC in mouse.

人类人工染色体（Human Artificial Chromosome，HAC）是一种具有来自于染色体chentromere区的重复DNA序列的染色体载体，它能与宿主染色体独立分离并稳定地保持在细胞内。最近，我们证明了HAC引入tetO序列（tetO-HAC）可以通过使用转录抑制子tTS诱导过量的异染色质而从细胞中消除。为了在小鼠中开发使用tetO-HAC的基因功能分析系统，我们产生了可以整合基因表达盒的tetO-HAC。然后，将GFP或tTS-EYFP表达盒整合到tetO-HAC中。在具有tTS-EYFP盒的tetO-HAC的情况下，在不存在Dox的情况下从细胞中消除tetO-HAC，这将异染色质诱导到tetO-HAC中。这些数据为建立tetO-HAC小鼠基因功能分析系统提供了重要依据。

项目成果

Generation of induced Pluripotent stem cells (iPSCs) using conditional human artificial chromosome (HAC)

使用条件人类人工染色体 (HAC) 生成诱导多能干细胞 (iPSC)

• 发表时间: 2012
• 作者: Yoshiaki Okamura;Tsukasa shimojima;Megumi Nakano;Jun-ichirou Ohzeki;Yoshinori Hasegawa;Masashi Ikeno;William C. Earnshaw;Vladimir Larionov and Hiroshi Masumoto
• 通讯作者: Vladimir Larionov and Hiroshi Masumoto