Tissue-Specific Transgenesis by Lentiviral Vectors

Abstract

Abstract: : Purpose: To develop a novel method to generate transgenic aminals using lentiviral vectors instead of conventional pronucleus injection. Using this method we produced several kinds of transgenic mice where heterologous genes were expressed in a tissue-specific manner in respect to the authentic genes. Methods: Lentiviral or retroviral vectors were coincubated with fertilized eggs for 48 hours after the removal of zona pellucida. Transduced blastcysts were implanted into uterus. We employed the upstream sequences of bovine rhodopsin gene, human red pigment gene and mouse keratin12 gene for the tissue-specific reporter gene expression in rod cells, cone cells and corneal epithelium, respectively. To evaluate transgenesis we performed PCR or Southern blot analyses. We confirmed tissue-specific reporter gene expression by histological and western blot analyses. Results: We obtained the transgenic mice using both lentiviral and retroviral vectors. The transgenes transduced by lentiviral vectors were not silenced but those transduced by retroviral vectors were silenced. We detected tissue-specific reporter gene expression in each transgenic mice using different tissue-specific promoters. Conclusions: Lentiviral vector is a useful tool for preparing transgenic mouse lines to identify cis-regulatory elements of tissue-specific promoters. Thereafter, tissue-specific promoters can be used for development of gene therapy strategy.

Keywords: gene transfer/gene therapy • photoreceptors • cornea: epithelium