کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1935614 | 1050670 | 2008 | 5 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: A lentiviral vector with novel multiple cloning sites: Stable transgene expression in vitro and in vivo A lentiviral vector with novel multiple cloning sites: Stable transgene expression in vitro and in vivo](/preview/png/1935614.png)
Gene delivery has become an important tool for biological research and gene therapy trials. Lentiviral vector (LV) mediated gene transfer is a preferred approach for stable, sustained transgenic expression. We report here step-wise development of an Indian human immunodeficiency virus type 2 (HIV-2) isolate derived third generation lentiviral vector with a novel, versatile multiple cloning site (MCS) that can also facilitate single step sub-cloning of a PCR amplified transgene cassette by T/A cloning strategy apart from useful cohesive/blunt end cloning. Efficiency of the vector systems was functionally demonstrated by development of a transgenic enhanced green fluorescence protein (GFP) expressing cell line. Further, a GFP down regulated cell line was derived from the said cell line through LV mediated shRNA expression by cloning the GFP-shRNA cassette using the T/A cloning strategy. Subsequently long term expression of GFP transgene in nude mouse spleen/liver was also documented till 30 days. This LV platform with the enhanced user friendly cloning options will be an important advancement in gene transfer technology.
Journal: Biochemical and Biophysical Research Communications - Volume 371, Issue 3, 4 July 2008, Pages 546–550