title: Site-specific transfer of an intact beta-globin gene cluster through a new targeting vector.
abstract: The ideal gene-therapy vector for treating genetic disorders should deliver intact therapeutic genes and their essential regulatory elements into the specific "safe genomic site" and realize long-term, self-regulatory expression. For beta-thalassemia gene therapy, viral vectors have been broadly used, but the accompanying insertional mutation and immunogenicity remain problematic. Hence, we aimed to develop new non-viral vectors that are efficient and safe in treating diseases. As previous studies have demonstrated that physiological expression of beta-globin genes requires both a 5' locus control region and 3' specific elements, we constructed a new human chromosome-derived targeting vector to transfer the intact beta-globin gene cluster into K562 cells. The whole beta-globin gene cluster was precisely integrated into the target site and expressed in a self-regulatory pattern. The results proved that the human chromosome-derived vector was specifically targeted to the human genome and this could provide a novel platform for further gene therapy research.
authors.affiliation: National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China.
authors.forename: Hai-sheng
authors.initials: HS
authors.lastname: Zhou
authors.forename: Na
authors.initials: N
authors.lastname: Zhao
authors.forename: Lei
authors.initials: L
authors.lastname: Li
authors.forename: Wen-ji
authors.initials: WJ
authors.lastname: Dong
authors.forename: Xue-song
authors.initials: XS
authors.lastname: Wu
authors.forename: De-long
authors.initials: DL
authors.lastname: Hao
authors.forename: Zhi-chen
authors.initials: ZC
authors.lastname: Guo
authors.forename: Kun
authors.initials: K
authors.lastname: Xia
authors.forename: Jia-hui
authors.initials: JH
authors.lastname: Xia
authors.forename: De-pei
authors.initials: DP
authors.lastname: Liu
authors.forename: Chih-chuan
authors.initials: CC
authors.lastname: Liang
mesh_terms: D001665:Binding Sites; D015139:Blotting, Southern; D015870:Gene Expression; D018390:Gene Targeting / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D005914:Globins / Q000235:genetics*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D020014:K562 Cells; D005810:Multigene Family; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods*
chemical_list: D012333:RNA, Messenger; D005914:Globins