生物
干细胞
病毒载体
基因表达
祖细胞
基因
基因表达谱
慢病毒
胚胎干细胞
基因治疗载体
重编程
报告基因
间充质干细胞
细胞生物学
分子生物学
遗传学
病毒
重组DNA
病毒性疾病
作者
Jun Tian,Stelios T. Andreadis
出处
期刊:Gene Therapy
[Springer Nature]
日期:2009-05-14
卷期号:16 (7): 874-884
被引量:57
摘要
Expression of multiple genes from the same target cell is required in several technological and therapeutic applications such as quantitative measurements of promoter activity or in vivo tracking of stem cells. In spite of such need, reaching independent and high-level dual-gene expression cannot be reliably accomplished by current gene transfer vehicles. To address this issue, we designed a lentiviral vector carrying two transcriptional units separated by polyadenylation, terminator and insulator sequences. With this design, the expression level of both genes was as high as that yielded from lentiviral vectors containing only a single transcriptional unit. Similar results were observed with several promoters and cell types including epidermal keratinocytes, bone marrow mesenchymal stem cells and hair follicle stem cells. Notably, we demonstrated quantitative dynamic monitoring of gene expression in primary cells with no need for selection protocols suggesting that this optimized lentivirus may be useful in high-throughput gene expression profiling studies.
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