Recombinase mediated cassette exchange into genomic targets using an adenovirus vector
Recombinase mediated cassette exchange into genomic targets using an adenovirus vector
复制标题
使用腺病毒载体将重组酶介导的盒式交换转化为基因组靶标
作者：
D. Sorrell;Claire J. Robinson;Jo;A. Kolb
重组酶介导的盒交换(RMCE)是一种位点特异性重组酶用一对不相容靶点的基因盒交换另一对相同位点的基因盒的过程。通常情况下，一个基因盒存在于宿主基因组中，而另一个基因盒通过化学或生物手段引入宿主细胞。我们在这里表明，盒式磁带交换的频率取决于转基因盒式磁带和重组酶的相对和绝对量。我们不仅通过电穿孔或化学介导的基因转移，而且通过携带要插入的转基因盒和重组酶编码区的腺病毒载体，成功地修饰了基因组靶点。RMCE在腺病毒载体能够复制的细胞中有效地进行。相比之下，在病毒不能复制的细胞中产生的转基因盒式磁带数量不足。额外的转基因盒式转染法显著提高了RMCE频率。这表明，在病毒载体的背景下，RMCE系统允许将转基因定向插入到定义的基因组位置。
Recombinase mediated cassette exchange (RMCE) is a process in which site-specific recombinases exchange one gene cassette flanked by a pair of incompatible target sites for another cassette flanked by an identical pair of sites. Typically one cassette is present in the host genome, whereas the other gene cassette is introduced into the host cell by chemical or biological means. We show here that the frequency of cassette exchange is dependent on the relative and absolute quantities of the transgene cassette and the recombinase. We were able to successfully modify genomic targets not only by electroporation or chemically mediated gene transfer but also by using an adenovirus vector carrying both the transgene cassette to be inserted and the recombinase coding region. RMCE proceeds efficiently in cells in which the adenovirus vector is able to replicate. In contrast, insufficient quantities of the transgene cassette are produced in cells in which the virus cannot replicate. Additional transfection of the transgene cassette significantly enhances the RMCE frequency. This demonstrates that an RMCE system in the context of a viral vector allows the site directed insertion of a transgene into a defined genomic site.
DOI：
10.1073/pnas.090527097
发表时间：
2000-05-23
影响因子：
11.1
作者：
Groth, AC;Olivares, EC;Calos, MP
通讯作者：
Calos, MP
影响因子：
20.3
作者：
Bouhassira, EE;Westerman, K;Leboulch, P
通讯作者：
Leboulch, P