Document Detail


Generation of HIV-1 based bi-cistronic lentiviral vectors for stable gene expression and live cell imaging.
MedLine Citation:
PMID:  23214259     Owner:  NLM     Status:  In-Process    
Abstract/OtherAbstract:
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.
Authors:
Lalit Sehgal; Srikanth Budnar; Khyati Bhatt; Sneha Sansare; Amitabha Mukhopadhaya; Rajiv D Kalraiya; Sorab N Dalal
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Indian journal of experimental biology     Volume:  50     ISSN:  0019-5189     ISO Abbreviation:  Indian J. Exp. Biol.     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-12-10     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0233411     Medline TA:  Indian J Exp Biol     Country:  India    
Other Details:
Languages:  eng     Pagination:  669-76     Citation Subset:  IM    
Affiliation:
KS215, Advanced Centre for Treatment Research & Education in Cancer, (ACTREC), Tata Memorial Centre, Kharghar Node, Navi Mumbai 410 210, India.
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