Polyadenylation signal facilitates the expression of foreign gene that is driven by an internal promoter located in the reverse orientation to long terminal repeat of retrovirus

We examined whether the presence of a polyadenylation [poly(A)] signal in a retrovirus vector could affect the expression level of exogenous gene(s) t hat was controlled by an internal promoter. Three suicide genes were placed under a promoter of the human midkine gene, whose expression is elevated i n lung cancer cells. Orientation of the internal transcriptional unit was d esigned to be opposite to the viral long terminal repeat. Expression of eac h suicide gene was greater in ecotropic packaging cells transfected with a retrovirus vector without a poly(A) signal than in those with a poly(A)-con taining vector. Sensitivity to ganciclovir, a prodrug that becomes an activ e drug by herpes simplex virus-thymidine kinase, was significantly improved in retrovirally transduced lung cancer cells compared with wild-type cells . However, the sensitivity was much greater in the cells transduced with po ly(A)-containing vector than in those with poly(A)-deleted construct. The p resence of a poly(A) signal downstream of exogenous gene(s) therefore favor s the expression of foreign gene(s) driven by an internal promoter.