Identification of a novel proliferation‐inducing determinant using lentiviral expression cloning
Chilov, Dmitri ; Fux, Cornelia ; Joch, Hana ; Fussenegger, Martin
In: Nucleic Acids Research, 2003, vol. 31, no. 18, p. e113-e113
Ajouter à la liste personnelle- 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