title: A highly stable and nonintegrated human artificial chromosome (HAC) containing the 2.4 Mb entire human dystrophin gene.
abstract: Episomal vector with the capacity to deliver a large gene containing all the critical regulatory elements is ideal for gene therapy. Human artificial chromosomes (HACs) have the capacity to deliver an extremely large genetic region to host cells without integration into the host genome, thus preventing possible insertional mutagenesis and genomic instability. Duchenne muscular dystrophy (DMD) is caused by mutation in the extremely large dystrophin gene (2.4 Mb). We herein report the development of a HAC vector containing the entire human dystrophin gene (DYS-HAC) that is stably maintained in mice and human immortalized mesenchymal stem cells (hiMSCs). The DYS-HAC was transferred to mouse embryonic stem (ES) cells, and isoforms of the DYS-HAC-derived human dystrophin in the chimeric mice generated from the ES cells were correctly expressed in tissue-specific manner. Thus, this HAC vector containing the entire dystrophin gene with its native regulatory elements is expected to be extremely useful for future gene and cell therapies of DMD.
authors.affiliation: Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University, Yonago, Japan.
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Hoshiya
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kazuki
authors.forename: Satoshi
authors.initials: S
authors.lastname: Abe
authors.forename: Masato
authors.initials: M
authors.lastname: Takiguchi
authors.forename: Naoyo
authors.initials: N
authors.lastname: Kajitani
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Watanabe
authors.forename: Toko
authors.initials: T
authors.lastname: Yoshino
authors.forename: Yasuaki
authors.initials: Y
authors.lastname: Shirayoshi
authors.forename: Katsumi
authors.initials: K
authors.lastname: Higaki
authors.forename: Graziella
authors.initials: G
authors.lastname: Messina
authors.forename: Giulio
authors.initials: G
authors.lastname: Cossu
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Oshimura
mesh_terms: D000818:Animals; D002460:Cell Line; D002645:Chickens; D022222:Chromosomes, Artificial, Human / Q000235:genetics*; D016189:Dystrophin / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D016189:Dystrophin