Site-specific transfer of an intact beta-globin gene cluster through a new targeting vector.
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.
Zhou
Hai-sheng
HS
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China.
Zhao
Na
N
Li
Lei
L
Dong
Wen-ji
WJ
Wu
Xue-song
XS
Hao
De-long
DL
Guo
Zhi-chen
ZC
Xia
Kun
K
Xia
Jia-hui
JH
Liu
De-pei
DP
Liang
Chih-chuan
CC
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*
D012333:RNA, Messenger; D005914:Globins