Development of gene functional analysis system using human artificial chromosome 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。

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.