Improvement of lentiviral vector-mediated gene transduction by genetic engineering of the structural protein Pr55Gag
Improvement of lentiviral vector-mediated gene transduction by genetic engineering of the structural protein Pr55Gag
复制标题
DOI：
10.1038/gt.2010.61
复制
发表时间：
2010-09-01
期刊：
影响因子：
5.1
通讯作者：
Komano, J.
中科院分区：
文献类型：
--
作者：
Aoki, T.;Shimizu, S.;Komano, J.
慢病毒载体是人类基因治疗的一种有前途的工具，因为它能够将基因转导到许多细胞类型中。然而，与慢病毒载体相关的技术问题之一是，与其他病毒载体相比，当前生产系统中的慢病毒滴度相对较低。在这项研究中，我们提供了遗传证据，表明在人类免疫缺陷病毒1型Pr55(Gag) (Gag)的氨基末端附着异源肉豆蔻酰化(myr)信号可以将病毒产量提高10倍，从而增强许多细胞系中的基因转导。 myr 信号 Gag 构建体在靶向耐去污剂膜区室、病毒出芽的 Vps4 依赖性以及病毒颗粒形态方面与野生型 Gag 表现相似。然而，通过活细胞中的生物发光共振能量转移测量，myr信号Gag构建体显示出提高的寡聚效率，有助于增加病毒产量和有效激活负责病毒颗粒成熟的病毒蛋白酶。转基因Gag代表了下一代慢病毒载体，应该有助于许多慢病毒载体应用的成功。基因治疗 (2010) 17, 1124-1133; doi：10.1038/gt.2010.61； 2010 年 4 月 22 日在线发布
The lentiviral vector is a promising tool for human gene therapy because of its ability to transduce genes into many cell types. However, one of the technical problems associated with the lentiviral vector is that lentiviral titers in current production systems are relatively low compared with the other viral vectors. In this study, we provide genetic evidence that the attachment of heterologous myristoylation (myr) signals on the amino-terminus of human immunodeficiency virus type 1 Pr55(Gag) (Gag) can increase the viral yield up to 10-fold, leading to the enhancement of gene transduction in many cell lines. The myr signal Gag constructs behaved similarly to the wild-type Gag in targeting to detergent-resistant membrane compartments, Vps4-dependence for viral budding, and virion morphology. However, the myr signal Gag constructs showed improved oligomerization efficiency as measured by bioluminescence resonance energy transfer in living cells, contributing to increased viral production and efficient activation of the viral protease responsible for virion maturation. The genetically modified Gag represents the next generation lentiviral vector, and should contribute to the success of many lentiviral vector applications. Gene Therapy (2010) 17, 1124-1133; doi:10.1038/gt.2010.61; published online 22 April 2010