Identification of a novel proliferation‐inducing determinant using lentiviral expression cloning

One of the major challenges in the post‐genome era is the correlation between genes and function or phenotype. We have pioneered a strategy for screening of cDNA libraries, which is based on sequential combination of lentiviral and oncoretroviral expression systems and can be used to identify prolif...

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Bibliographic Details
Published in:Nucleic acids research 2003-09, Vol.31 (18), p.e113-e113
Main Authors: Chilov, Dmitri, Fux, Cornelia, Joch, Hana, Fussenegger, Martin
Format: Article
Language:English
Subjects:
3T3 Cells
Adult
Animals
Brain - metabolism
Cell Division - genetics
Cell Line
Cells, Cultured
Cloning, Molecular - methods
DNA, Complementary - chemistry
DNA, Complementary - genetics
Flow Cytometry
Gene Expression
Gene Library
Genetic Vectors - genetics
Green Fluorescent Proteins
HeLa Cells
Humans
Lentivirus - genetics
Luminescent Proteins - genetics
Luminescent Proteins - metabolism
Mice
Molecular Sequence Data
NAR Methods Online
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Sequence Analysis, DNA
Transfection
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Summary:One of the major challenges in the post‐genome era is the correlation between genes and function or phenotype. We have pioneered a strategy for screening of cDNA libraries, which is based on sequential combination of lentiviral and oncoretroviral expression systems and can be used to identify proliferation‐modulating genes. Screening of a lentiviral expression library derived from adult human brain cDNA resulted in cloning of the potent proliferation‐inducing determinant termed pi1 (proliferation inducer 1). Transduction experiments using GFP‐expressing oncoretroviruses to target proliferation‐competent cells suggested that overexpression of pi1 initiates proliferation of human umbilical vein endothelial cells (HUVECs). Growth induction of HUVECs as well as Swiss3T3 fibroblasts was confirmed by Brd‐uridine incorporation assays, which correlated increased DNA synthesis with expression of pi1. The identified pi1 cDNA is 297 bp long and encodes a 10 kDa polypeptide. Since deregulation of proliferation control accounts for a number of today’s untreatable human diseases such as neurodegenerative disorders and cancer, discovery of novel proliferation‐modulating genes is essential for developing new strategies for gene therapy and tissue engineering.
ISSN:0305-1048
1362-4962
1362-4962
DOI:10.1093/nar/gng115