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IJEB Vol.50 [2012] >
IJEB Vol.50(10) [October 2012] >
| Title: | Generation of HIV-1 based bi-cistronic lentiviral vectors for stable gene expression and live cell imaging. |
| Authors: | Sehgal, Lalit
Budnar, Srikanth
Bhatt, Khyati
Sansare, Sneha
Mukhopadhaya, Amitabha
Kalraiya, Rajiv D
Dalal, Sorab N |
| Keywords: | Bi-cistronic lentiviral vector
FRET
Fluorescent tag
HIV-I
In vivo imaging
Multiple cloning site
Stable transgene expression |
| Issue Date: | Oct-2012 |
| Publisher: | NISCAIR-CSIR, India |
| Abstract: | The study of protein-protein interactions, protein
localization, protein organization into higher order structures and organelle
dynamics in live cells, has greatly enhanced the understanding of various
cellular processes. Live cell imaging
experiments employ plasmid or viral vectors to express the protein/proteins of
interest fused to a fluorescent protein. Unlike plasmid vectors, lentiviral
vectors can be introduced into both dividing and non dividing cells, can be
pseudotyped to infect a broad or narrow range of cells, and can be used to
generate transgenic animals. However, the currently available lentiviral
vectors are limited by the choice of fluorescent protein tag, choice of
restriction enzyme sites in the Multiple Cloning Sites (MCS) and promoter
choice for gene expression. In this report, HIV-1 based bi-cistronic lentiviral
vectors have been generated that drive the expression of multiple fluorescent
tags (EGFP, mCherry, ECFP, EYFP and dsRed), using two different promoters. The presence of a unique MCS with multiple
restriction sites allows the generation of fusion proteins with the fluorescent
tag of choice, allowing analysis of multiple fusion proteins in live cell
imaging experiments. These novel lentiviral vectors are improved delivery
vehicles for gene transfer applications and are important tools for live cell
imaging in vivo.
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| Page(s): | 669-676 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-1009 (Online); 0019-5189 (Print) |
| Source: | IJEB Vol.50(10) [October 2012]
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