Human Artificial Chromosome (Hac) Vector With A Conditional Centromere For Correction Of Human Genetic Deficiencies

Human artificial chromosome (HAC)-based vectors offer a promising system for delivery and expression of full-length human genes of any size. Due to their unique maintenance features and unlimited cloning capacity, HACs avoid the lack of copy number control and insertional mutagenesis caused by integration into host chromosomes that plague viral vectors. We previously described a synthetic HAC that can be easily eliminated from cell populations by inactivation of its conditional kinetochore. Here, we demonstrate the utility of this HAC, which has a unique gene acceptor site, for delivery of full-length genes and correction of genetic deficiencies in human cells. A battery of functional tests was performed to demonstrate expression of NBS1 and VHL genes from the HAC at physiological levels. We also show that phenotypes arising from stable gene expression can be reversed when cells are “cured” of the HAC by inactivating its kinetochore in proliferating cell populations, a feature that provides a control for phenotypic changes attributed to expression of HAC-encoded genes. This generation of human artificial chromosomes should be suitable for studies of gene function and therapeutic applications. Citation Format: Jung-Hyun Kim, Artem Kononenko, Indri Erliandri, Tae-Aug Kim, Megumi Nakano, Yuichi Iida, J. Carl Barrett, Mitsuo Oshimura, Hiroshi Masumoto, William C. Earnshaw, Vladimir Larionov, Natalay Kouprina. Human artificial chromosome (HAC) vector with a conditional centromere for correction of human genetic deficiencies. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3310. doi:10.1158/1538-7445.AM2013-3310