A dicistronic construct allows easy detection of human CFTR expression from YAC DNA in human cells

Abstract

We have made a dicistronic construct where the picornaviral internal ribosome-entry site (IRES) driving the expression of the β-geo gene has been inserted into the 3' untranslated region of the human CFTR gene present in a YAC. When introduced into the human cell line Caco-2 expressing the CFTR gene, the expression of the dicistronic gene can be detected by lacZ staining and follows the accumulation of the endogenous CFTR mRNA upon differentiation of the cells. These data demonstrate that this IRES-based approach presents an alternative to mRNA in situ hybridisation and allows detection of expression in an autologous system.