Generation of an improved foamy virus vector by dissection of cis-acting sequences.
Generation of an improved foamy virus vector by dissection of cis-acting sequences.
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通过解剖顺式作用序列生成改进的泡沫病毒载体。
DOI：
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发表时间：
2009
影响因子：
3.8
通讯作者：
A. Rethwilm
中科院分区：
文献类型：
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作者：
T. Wiktorowicz;Katrin Peters;Nicole S. Armbruster;A. Steinert;A. Rethwilm
与其他逆转录病毒不同，泡沫病毒(FV)从剪接的mRNA中独立地产生Pol蛋白前体，而不是Gag蛋白。POL蛋白进入病毒衣壳的确切机制尚不清楚。在此之前，我们已经证明POL的囊化关键依赖于(前)基因组RNA的包装，并在Pol衣壳掺入所需的顺式作用序列(CASI和CASII)中发现了两个不同的信号，即POL囊化序列(PESI和PESII)。在这里，我们调查了FV载体中PESI和PESII的存在是否足以包裹Pol，以及是否可以在不损失功能的情况下缩短相当扩展的CASII元件。我们的结果表明：(I)Pesi和II的存在不足以包裹Pol，(Ii)带有缩短的CASII元件的原型Fv载体保留了Pol的掺入和完整的功能，特别是在转导成纤维细胞和原代人间充质干细胞时，(Iii)中央多嘌呤通路的存在显著提高了Fv载体的转导效率，(Iv)Pol包裹和RNA包装可以明显分离。本质上，我们设计了一个新的FV载体，它比以前建立的载体少了大约850个核苷酸的CAS，并且在分析转导细胞系和原代人类细胞时完全具有功能。
In contrast to other retroviruses, foamy viruses (FVs) generate their Pol protein precursor independently of the Gag protein from a spliced mRNA. The exact mechanism of Pol protein incorporation into the viral capsid is poorly understood. Previously, we showed that Pol encapsidation critically depends on the packaging of (pre-) genomic RNA and identified two distinct signals within the cis-acting sequences (CASI and CASII), Pol encapsidation sequences (PESI and PESII), which are required for Pol capsid incorporation. Here, we investigated whether the presence of PESI and PESII in an FV vector is sufficient for Pol encapsidation and whether the rather extended CASII element can be shortened without loss of functionality. Our results indicate that (i) the presence of PESI and II are not sufficient for Pol encapsidation, (ii) prototype FV vectors with a shortened CASII element retain Pol incorporation and full functionality, in particular upon transducing fibroblasts and primary human mesenchymal stem cells, (iii) the presence of the central poly purine tract significantly increased the transduction rates of FV vectors and (iv) Pol encapsidation and RNA packaging can be clearly separated. In essence, we designed a new FV vector that bears approximately 850 bp less of CAS than previously established vectors and is fully functional when analysed to transduce cell lines and primary human cells.
影响因子：
12.4
作者：
Trobridge, G;Josephson, N;Russell, DW
通讯作者：
Russell, DW
影响因子：
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作者：
Nöth, U;Tuli, R;Tuan, RS
通讯作者：
Tuan, RS