Efficient transduction of myeloid cells by an HIV-1-derived lentiviral vector that packages the Vpx accessory protein.
Efficient transduction of myeloid cells by an HIV-1-derived lentiviral vector that packages the Vpx accessory protein.
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DOI：
10.1038/gt.2012.61
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发表时间：
2013-05
期刊：
影响因子：
5.1
通讯作者：
Landau, N. R.
中科院分区：
文献类型：
--
作者：
Bobadilla, S.;Sunseri, N.;Landau, N. R.
慢病毒载体被广泛用于基因的稳定表达和shrna介导的敲低，目前正在开发用于基因治疗的临床应用。VSV-G的假分型使它们能够感染广泛的细胞类型。然而，骨髓细胞，如树突状细胞和巨噬细胞，由于骨髓特异性限制因子SAMHD1的存在，对慢病毒载体转导相对难耐受。SIVmac/HIV-2和相关病毒通过编码Vpx来缓解SAMHD1介导的限制，Vpx是一种病毒粒子包装的辅助蛋白，在感染时诱导SAMHD1降解。HIV-1不编码Vpx，也不能包装这种蛋白质。我们报道了一种基于hiv -1的慢病毒载体的开发，其中Vpx包装基序被放置在Gag/Pol表达载体的p6区域，用于生成慢病毒载体病毒粒子。病毒粒子以高拷贝数包装Vpx，并以2倍的滴度增加感染髓细胞。树突状细胞与转运蛋白-3的shRNA转导导致编码mRNA的90%被敲除。该系统可应用于任何基于hiv的慢病毒载体，在需要骨髓细胞有效转导的实验室和临床应用中非常有用。
Lentiviral vectors are widely used for the stable expression of genes and shRNA-mediated knockdown and are currently under development for clinical use in gene therapy. Pseudotyping of the vectors with VSV-G allows them to infect a wide range of cell-types. However, myeloid cells, such as dendritic cells and macrophages are relatively refractory to lentiviral vector transduction as a result of the myeloid-specific restriction factor, SAMHD1. SIVmac/HIV-2 and related viruses relieve the SAMHD1-mediated restriction by encoding Vpx, a virion-packaged accessory protein that induces the degradation of SAMHD1 upon infection. HIV-1 does not encode Vpx and cannot package the protein. We report the development of an HIV-1-based lentiviral vector in which the Vpx packaging motif has been placed in the p6 region of the Gag/Pol expression vector that is used to generate the lentiviral vector virions. The virions package Vpx in high copy number and infect myeloid cells with a two-log increase in titer. Transduction of dentritic cells with an shRNA against transportin-3 resulted in >90% knock-down of the encoding mRNA. The system can be applied to any HIV-based lentiviral vector and is useful for laboratory and clinical applications where the efficient transduction of myeloid cells is required.
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影响因子：
64.8
作者：
通讯作者：
--
DOI：
10.1016/s0167-4781(99)00046-9
发表时间：
1999-05-14
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION
影响因子：
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作者：
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作者：
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作者：
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影响因子：
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作者：
Goldstone, David C.;Ennis-Adeniran, Valerie;Webb, Michelle
通讯作者：
Webb, Michelle