Fig 1. Schematic illustration of the principle of the RINES system.
A) Representation of the RINES plasmid construct with localization of the miRNA targeting sequence (miR T) and the hNIS cDNA reporter gene. B) Left panel, when the miRNA of interest is not expressed in cells, the CymR mRNA is transcribed as well as the CymR protein which binds to the Cuo operator sequence located in the CMV5/Cuo inducible expression cassette. Under this configuration, hNIS expression is switched-OFF and there is no, or low 99mTc04- accumulation in cells. Right panel, in contrast when present in cells, the miRNA of interest binds to the miR T sequence located in the 3′-UTR of the CymR repressor transcript, inhibiting production of the CymR protein through activation of the RISC machinery. Under this configuration, hNIS is switched-ON, resulting in expression of hNIS at the cellular membrane and accumulation of 99mTc04- in cells.