Project Details
Projekt
Gene therapy with designed splice factors to correct splicing defects in Rd6 mice
Applicant
Professor Dr. John Neidhardt
Subject Area
Molecular Biology and Physiology of Neurons and Glial Cells
Term
from 2018 to 2022
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 399447659
Approximately 10-20% of all mutations causing genetic diseases interfere with splicing. With few exceptions, this observation is made irrespective of the disease gene. Consequently, gene therapeutic approaches that correct splice defects are highly attractive treatment options for many inherited diseases, including retinal degeneration. As the eye constitutes an ideal target organ for gene therapeutic interventions, we developed gene therapeutic approaches that are efficient in correcting splice donor site defects focusing on genes associated with retinal degeneration.Our therapies apply designed U1- and U6-snRNA-based splice factors that ameliorate the effect of splice donor site mutations. The functionality and efficacy of the approach was successfully tested in several different patient-derived cell lines on RNA and protein level. Furthermore, the preliminary results presented in this proposal strongly suggest that the UsnRNA-based therapeutic approach is efficient in retinal cells after subretinal injection and AAV-mediated transduction in vivo.We aim to treat the Rd6 mouse model which carries a splice donor site mutation in the Mfrp gene. The mutation in the Rd6 mouse causes a splice defect (skipping of Mfrp exon 4) and leads to retinitis pigmentosa-like retinal degeneration and flecked retinal appearance. The proposed project will evaluate and optimize gene therapeutic treatments of the Rd6 mouse model applying designed U1- and U6-snRNAs to correct the mutation-induced splice defect in the retina and in vivo. We will evaluate the efficacy of AAV-mediated therapies in the retina of mutant and control Rd6 mice using histological, molecular and functional testing. In addition, we include safety measures into the evaluation of our therapeutic approach.The experiments proposed in this application will help to establish RNA-based treatments suitable not only in the Rd6 context, but also for several other monogenetic diseases of the retina. The project is well-embedded in the SPP2127 program and will further guide the way to develop highly promising gene therapies for large animal models or human patients.
DFG Programme
Priority Programmes
