A non-viral vector for potential DMD gene therapy study by targeting a minidystrophin-GFP fusion gene into the hrDNA locus.
Gene therapy has emerged as a promising approach for the lethal disorder of Duchenne muscular dystrophy (DMD). Using a novel non-viral delivery system, the human ribosomal DNA (hrDNA) targeting vector, we targeted a minidystrophin-GFP fusion gene into the hrDNA locus of HT1080 cells with a high site-specific integrated efficiency of 10(-5), in which the transgene could express efficiently and continuously. The minidystrophin-GFP fusion protein was easily found to localize on the plasma membrane of HT1080 cells, indicating its possible physiologic performance. Our findings showed that the hrDNA-targeting vector might be highly useful for DMD gene therapy study.
Yang
Junlin
J
State Key Laboratory of Medical Genetics, Central South University, Changsha, China.
Liu
Xionghao
X
Yu
Jiaoling
J
Sheng
Liang
L
Shi
Yan
Y
Li
Zhuo
Z
Hu
Youjin
Y
Xue
Jinfeng
J
Wu
Lingqian
L
Liang
Yu
Y
Xia
Jiahui
J
Liang
Desheng
D
D015153:Blotting, Western; D004275:DNA, Ribosomal / Q000235:genetics*; D016189:Dystrophin / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D018390:Gene Targeting; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics* / Q000378:metabolism / Q000628:therapy*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019076:Transgenes / Q000235:genetics; D014407:Tumor Cells, Cultured
D004275:DNA, Ribosomal; D016189:Dystrophin; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins