Silencing of the cytosolic 5'nucleotidase II (cN-II)mediated by a shRNA-expressing lentiviral vector.

Cytosolic 5’-nucleotidase/phosphotransferase (cN-II), acting through the formation of a phospho-enzyme intermediate, is specific for oxy- and deoxypurine monophosphates. cN-II is an enzyme of great importance: overexpression of cN-II has negative effects on human cell survival. cN-II is involved in the regulation of intracellular concentration of IMP, AMP, GMP and also PRPP, therefore in the regulation of purine de novo synthesis and salvage. cN-II is clinically interesting, since it is involved in prodrug cellular metabolism. Besides, mRNA level of cN-II has a prognostic value in adult AML (acute myeloid leukemia). From this view point, the study of cN-II depletion constitutes a crucial step in the understanding of its functions in cell metabolism. The knock-down of cN-II revealed that its absence is lethal for cells, and that the addition of the product normally generated by the enzyme allows knocked-down cells to survive.