Department of Animal Science, Iowa State University, Ames, Iowa, USA.
Invest Ophthalmol Vis Sci. 2012 Jan 31;53(1):501-7. doi: 10.1167/iovs.11-8784.
The Pro23His (P23H) rhodopsin (RHO) mutation underlies the most common form of human autosomal dominant retinitis pigmentosa (adRP). The objective of this investigation was to establish a transgenic miniature swine model of RP using the human P23H RHO gene.
Somatic cell nuclear transfer (SCNT) was used to create transgenic miniature pigs that expressed the human P23H RHO mutation. From these experiments, six transgenic founders were identified whose retinal function was studied with full-field electroretinography (ffERG) from 3 months through 2 years. Progeny from one founder were generated and genotyped to determine transgene inheritance pattern. Retinal mRNA was isolated, and the ratio of P23H to wild-type pig RHO was measured.
A single transgene integration site was observed for five of the six founders. All founders had abnormal scotopic and photopic ffERGs after 3 months. The severity of the ffERG phenotype was grouped into moderately and severely affected groups. Offspring of one founder inherited the transgene as an autosomal dominant mutation. mRNA analyses demonstrated that approximately 80% of total RHO was mutant P23H.
Expression of the human RHO P23H transgene in the retina creates a miniature swine model with an inheritance pattern and retinal function that mimics adRP. This large-animal model can serve as a novel tool for the study of the pathogenesis and therapeutic intervention in the most common form of adRP.
Pro23His（P23H）视蛋白（RHO）突变是人类常染色体显性遗传视网膜色素变性（adRP）最常见的形式。本研究的目的是利用人 P23H RHO 基因突变建立一个 RP 的转基因小型猪模型。
体细胞核移植（SCNT）用于创建表达人 P23H RHO 突变的转基因小型猪。从这些实验中，确定了 6 个转基因创始者，其视网膜功能通过全视野视网膜电图（ffERG）从 3 个月到 2 年进行研究。从一个创始者中产生了后代，并对其进行了基因分型以确定转基因的遗传模式。分离视网膜 mRNA，并测量 P23H 与野生型猪 RHO 的比例。
观察到 6 个创始者中的 5 个有一个单一的转基因整合位点。所有创始者在 3 个月后都出现了异常的暗视野和明视野 ffERG。ffERG 表型的严重程度分为中度和重度影响组。一个创始者的后代遗传了该转基因作为常染色体显性突变。mRNA 分析表明，约 80%的总 RHO 是突变的 P23H。
视网膜中表达人 RHO P23H 转基因可创建一种与 adRP 遗传模式和视网膜功能相似的小型猪模型。这种大型动物模型可以作为研究最常见形式的 adRP 的发病机制和治疗干预的新工具。
