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非整合性泡沫病毒载体。

Nonintegrating foamy virus vectors.

机构信息

Department of Medicine, University of Washington, 1959 NE Pacific St., Seattle, WA 98195, USA.

出版信息

J Virol. 2010 Sep;84(18):9341-9. doi: 10.1128/JVI.00394-10. Epub 2010 Jun 30.

Abstract

Foamy viruses (FVs), or spumaviruses, are integrating retroviruses that have been developed as vectors. Here we generated nonintegrating foamy virus (NIFV) vectors by introducing point mutations into the highly conserved DD35E catalytic core motif of the foamy virus integrase sequence. NIFV vectors produced high-titer stocks, transduced dividing cells, and did not integrate. Cells infected with NIFV vectors contained episomal vector genomes that consisted of linear, 1-long-terminal-repeat (1-LTR), and 2-LTR circular DNAs. These episomes expressed transgenes, were stable, and became progressively diluted in the dividing cell population. 1-LTR circles but not 2-LTR circles were found in all vector stocks prior to infection. Residual integration of NIFV vectors occurred at a frequency 4 logs lower than that of integrase-proficient FV vectors. Cre recombinase expressed from a NIFV vector mediated excision of both an integrated, floxed FV vector and a gene-targeted neo expression cassette, demonstrating the utility of these episomal vectors. The broad host range and large packaging capacity of NIFV vectors should make them useful for a variety of applications requiring transient gene expression.

摘要

泡沫病毒(FV),也称为泡沫状病毒,是整合性逆转录病毒,已被开发为载体。在这里,我们通过在泡沫状病毒整合酶序列的高度保守的 DD35E 催化核心基序中引入点突变,产生了非整合性泡沫病毒(NIFV)载体。NIFV 载体产生高滴度的病毒 stocks,转导分裂细胞,并且不整合。感染 NIFV 载体的细胞含有由线性、1 个长末端重复(1-LTR)和 2 个 LTR 环状 DNA 组成的游离载体基因组。这些游离体表达转基因,稳定,并且在分裂细胞群中逐渐稀释。在感染之前,所有载体 stocks 中都发现了 1-LTR 环,但没有 2-LTR 环。NIFV 载体的残余整合频率比整合酶有效的 FV 载体低 4 个对数级。来自 NIFV 载体的 Cre 重组酶介导了整合的、floxed FV 载体和基因靶向 neo 表达盒的切除,证明了这些游离载体的实用性。NIFV 载体的广泛宿主范围和大容量包装能力应该使它们在需要瞬时基因表达的各种应用中非常有用。

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