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Yuichi Iida, Yasuhiro Kazuki and Mitsuo Oshimura
Posters: Adv Genet Eng
Conventional gene delivery vectors such as viral vectors, plasmid, bacterial artificial chromosome (BAC) might be associated with transgene-silencing because of position effect. Human artificial chromosome (HAC) vector is considered as a promising system for gene delivery and express. The HAC is produced by top-down and bottom-up approaches. Both HAC can be episomally maintained in their nuclei and segregate normally. The top-down approach can generate linear HAC ranging in approximately 3~4 Mb that retain natural centromeres and are mitotically stable in human and mouse cells. Moreover, we have succeeded in making trans-chromosomic-mice containing a HAC. It can be maintained in vivo and can produce mouse model. The bottom-up approach of HAC generation was initially developed in human fibrosarcoma HT1080 cells by introducing synthetic alphoid DNA cloned into a circular BAC vector. The tetO-HAC, generated by bottom-up approach, can be eliminated from mammalian cells such as CHO and HT1080 cells. The tetO-HAC containing human chromosome 17 alphoid DNA and tet-operater sequences in its centromeric regions resulted in controlling the centromere function. We have demonstrated that the tetO-HAC can be eliminated from nuclei in human HT1080 remarkably. The combination of using two types of HAC would be a powerful tool for cell reprogramming and gene collection of genetic disorders.
Yuichi Iida has completed his masters degree at the age of 23 years from Tottori University, and is currently a graduate student studying for his doctoral thesis. He has published 5 papers in reputed journals