Hyperactive transposase mutants of the Sleeping Beauty transposon

被引:78
作者
Baus, J
Liu, L
Heggestad, AD
Sanz, S
Fletcher, BS
机构
[1] Univ Florida, Coll Med, Dept Pharm & Therapeut, Gainesville, FL 32610 USA
[2] Vet Affairs Med Ctr, Med Res Serv, Gainesville, FL 32608 USA
关键词
Sleeping Beauty; transposon; gene therapy; nonviral vector; polyethylenimine;
D O I
10.1016/j.ymthe.2005.06.484
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Transposable elements have enormous potential to overcome one of the major hurdles in nonviral gene delivery, namely the lack of long-term gene expression. The Sleeping Beauty (SB) transposon is a promising vector system for nonviral gene therapy as it has the highest transposition activity of all known DNA transposons within mammalian cells. In an effort to generate a more efficient delivery vehicle, we conducted a systematic evaluation of several novel and previously identified SB transposase mutants. The results indicate that certain combinations of mutants do not enhance transposition, whereas others give a synergistic response. The most active mutant, designated HSB17, shows nearly 17-fold higher transposition activity compared to the original transposase SB10 when tested within the same expression cassette. In addition, synergistic activity is observed when this hyperactive mutant is combined with an improved transposon. Animal studies utilizing the hyperactive transposase show enhanced long-term reporter gene expression. These modifications further expand the utility of this transposon-based gene transfer system.
引用
收藏
页码:1148 / 1156
页数:9
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