Abstract
This chapter discusses a transient expression system that is suitable for the study of the expression of the interstitial retinoid-binding protein (IRBP) gene. This system is applicable for gene expression studies of photoreceptor-specific proteins in retinoblastoma cells. A retinoblastoma cell line, WERI-RB1, is used to dissect the regulatory sequences that are responsible for the expression of human IRBP in vivo . IRBP is a large and elongated glycoprotein that is universally distributed throughout the vertebrates. It is synthesized by the photoreceptor cells in the retina and is secreted into the interphotoreceptor matrix. The major function of IRBP is to transport 11-cis- and all-trans-retinoids between the neural retina and the retinal pigment epithelium. The chapter discusses the optimization of WERI-RB1 transfections. The conditions required for optimal transfection vary depending on each individual cell line. The amount of the plasmid construct and the liposome used for transfection should be adjusted so that the transfected cells produce maximum chloramphenicol acetyltransferase (CAT) enzyme activity for a given promoter.
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