Deoxyribonuclease II is a lysosomal barrier to transfection

被引：64

作者：

Howell, DPG

Krieser, RJ

Eastman, A

Barry, MA ^{[1
]}

机构：

[1] Baylor Coll Med, Ctr Cell & Gene Therapy, Houston, TX 77030 USA

[2] Baylor Coll Med, Dept Microbiol & Immunol, Houston, TX 77030 USA

[3] Baylor Coll Med, Dept Mol & Human Genet, Houston, TX 77030 USA

[4] Dartmouth Coll Sch Med, Dept Pharm & Therapeut, Hanover, NH 03755 USA

[5] Rice Univ, Dept Bioengn, Houston, TX 77005 USA

来源：

MOLECULAR THERAPY | 2003年 / 8卷 / 06期

关键词：

D O I：

10.1016/j.ymthe.2003.09.011

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：

DNA delivered in nonviral vectors or as naked DNA must overcome a number of extracellular and intracellular barriers to transfection. Since many vectors deliver DNA into cells by the endocytic route, DNA degradation by lysosomal nucleases has been proposed as a significant barrier to transfection, despite the fact that this has not yet been formally demonstrated to occur. To test this hypothesis, we have investigated the role of deoxyribonuclease II (DNase II), the primary acidic endonuclease active in the lysosome, in transfection. Two genetic systems were engineered in which mammalian cells either overexpressed DNase II or were knocked out for the enzyme. In both models, higher levels of DNase II correlated with decreased transfection efficiency by nonviral DNA delivery vectors. These data provide direct evidence implicating lysosomal DNase II as a barrier to transfection.

引用

页码：957 / 963

页数：7

共 30 条

[1] Molecular cloning and characterization of human and murine DNase II [J].