'Advanced' generation lentivirruses as efficient vectors for cardiomyocyte gene transduction in vitro and in vivo
'Advanced' generation lentivirruses as efficient vectors for cardiomyocyte gene transduction in vitro and in vivo
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DOI：
10.1038/sj.gt.3301936
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发表时间：
2003-04-01
期刊：
影响因子：
5.1
通讯作者：
Condorelli, G
中科院分区：
文献类型：
--
作者：
Bonci, D;Cittadini, A;Condorelli, G
心肌细胞中的有效基因转导是一项只能通过病毒载体完成的任务。到目前为止，用于此目的的最常用载体是腺病毒衍生的载体。最近，已经证明慢病毒载体可以抑制生长停滞的细胞，如造血干细胞。此外，与第三代载体相比，含有mRNA稳定序列和核输入序列的修饰形式的慢病毒载体（“高级”一代）已被证明可以显着改善生长停滞细胞中的基因转导。因此，我们测试了“高级”代慢病毒是否能够在体外和体内感染和转导心肌细胞，并与第三代相同载体的体外功效进行比较。在这里，我们报告说，“先进”的一代慢病毒载体感染大多数（> 80%）心肌细胞的培养，如免疫荧光和流式细胞仪分析所示：相比之下，心肌细胞感染的第三代慢病毒的百分比低三至四倍。此外，“高级”代慢病毒也能够在体内感染成人心肌并诱导稳定的基因表达。因此，“先进”一代慢病毒载体可用于心肌细胞中的体外和体内基因表达研究。
Efficient gene transduction in cardiomyocytes is a task that can be accomplished only by viral vectors. Up to now, the most commonly used vectors for this purpose have been adenoviral-derived ones. Recently, it has been demonstrated that lentiviral vectors can transduce growth-arrested cells, such as hematopoietic stem cells. Moreover, a modified form of lentiviral vector (the 'advanced' generation), containing an mRNA-stabilizer sequence and a nuclear import sequence, has been shown to significantly improve gene transduction in growth-arrested cells as compared to the third-generation vector. Therefore, we tested whether the 'advanced' generation lentivirus is capable of infecting and transducing cardiomyocytes both in vitro and in vivo, comparing efficacy in vitro against the third-generation of the same vector. Here we report that 'advanced' generation lentiviral vectors infected most (> 80%) cardiomyocytes in culture, as demonstrated by immunofluorescence and FACS analyses: in contrast the percentage of cardiomyocytes infected by third-generation lentivirus was three- to four-fold lower. Moreover, 'advanced' generation lentivirus was also capable of infecting and inducing stable gene expression in adult myocardium in vivo. Thus, 'advanced' generation lentiviral vectors can be used for both in vitro and in vivo gene expression studies in the cardiomyocyte.