Nature Biotechnology
18, 1086 - 1090 (2000)
doi:10.1038/80287
Manipulation of human minichromosomes to carry greater than megabase-sized chromosome insertsYoshimi Kuroiwa1, 3, Kazuma Tomizuka1, 3, Tokuyuki Shinohara2, Yasuhiro Kazuki2, Hitoshi Yoshida1, Atsuko Ohguma1, Terumi Yamamoto1, Sonoko Tanaka1, Mitsuo Oshimura2
& Isao Ishida11
Pharmaceutical Research Laboratory, Kirin Brewery. Co., Ltd., 3 Miyahara-cho Takasaki-shi Gunma 370-1295, Japan. 2
Department of Molecular and Cell Genetics, School of Life Sciences, Faculty of Medicine, Tottori University, Nishimachi 86 Yonago, Tottori 683-8503, Japan. 3
These authors contributed equally to this work.
Correspondence should be addressed to Isao Ishida i-ishida@kirin.co.jpchromosome cloninghuman artificial chromosomesFor introducing regions of human chromosomes greater than a megabase into cells or animals, we have developed a chromosome-cloning system in which defined regions of human chromosomes can be cloned into a stable human minichromosome vector in homologous recombination-proficient chicken DT40 cells. The stable minichromosome vector allowed a 10 Mb-sized region of the mitotically unstable human chromosome 22 to be stably maintained in mouse embryonic stem (ES) cells, and in mice. Furthermore, we demonstrated functional expression of human genes from the HAC in mice. This study describes a stable cloning and expression system for greater than megabase−sized regions of human chromosomes.
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