Retina-specific gene excision by targeted expression of Cre recombinase

Abstract

The use of Cre recombinase for conditional targeting permits the controlled removal or activation of genes in specific tissues and at specific times of development. The Rho-Cre mice provide an improved tool for studying gene ablation in rod photoreceptor cells. To establish a robust expression of Rho-Cre transgenic mice that would be useful for the study of various protein functions in photoreceptor cells, a total 11,987 kb fragment (pNCHS4 Rho-NLS-cre) containing human rhodopsin promoter was cloned. The Rho-Cre plasmid was digested with EcoR1 and I Ceu-1, and the 9.316 kb fragment containing the hRho promoter and Cre recombinase gel was purified. To generate transgenic mice, the purified DNA fragment was injected into fertilized oocytes according to standard protocols. ROSA26R reported the steady expression of Rho-Cre especially in photoreceptor cells, allowing further excising proteins in rod photoreceptors across the retina. This Rho-Cre transgenic line should thus prove useful as a general deletor line for genetic analysis of diverse aspects of retinopathy.

Keywords: Cre recombinase; EGTA; OCT; PBS; PCR; PFA; Retina; Rho; SPF; Transgenic mice; ethyleneglycol bis(2-aminoethylether)tetraacetic acid; opti-mum cutting temperature compound; paraformaldehyde; phosphate buffer saline; polymerase chain reaction; specific-pathogen-free.