A5065764993- Retinal Development and Disorders
- Virus-based gene therapy research
- CRISPR and Genetic Engineering
- interferon and immune responses
- Genetic and Kidney Cyst Diseases
- Retinopathy of Prematurity Studies
- Herpesvirus Infections and Treatments
- Retinal and Optic Conditions
- Retinal Diseases and Treatments
- Vascular Malformations and Hemangiomas
- Cytomegalovirus and herpesvirus research
- Congenital Ear and Nasal Anomalies
- Neuroscience and Neural Engineering
Telethon Institute Of Genetics And Medicine
2015-2022
Adeno-associated viral vectors reconstitute large proteins in the retina via intein-mediated protein trans-splicing.
Retinal gene transfer with adeno-associated viral (AAV) vectors holds great promise for the treatment of inherited retinal degenerations (IRDs). One limit AAV is its capacity about 5 kb, which can be expanded to 9 using dual vectors. This strategy would still not suffice IRDs such as Usher syndrome type 1D or Alström I (ALMS) due mutations in CDH23 ALMS1, respectively. To overcome this limitation, we generated triple vectors, a maximal 14 kb. Transcriptomic analysis following transduction...
Abstract Challenges to the widespread application of gene therapy with adeno-associated viral (AAV) vectors include dominant conditions due gain-of-function mutations which require allele-specific knockout, as well long-term transgene expression from proliferating tissues, is hampered by AAV DNA episomal status. To overcome these challenges, we used CRISPR/Cas9-mediated homology-independent targeted integration (HITI) in retina and liver paradigmatic target tissues. We show that AAV-HITI...
Split intein-mediated protein trans-splicing expands AAV transfer capacity, thus overcoming the limited cargo. However, non-mammalian inteins persist as by-products, and this could raise safety concerns for intein clinical applications. In study, we tested ability of several degrons to selectively decrease levels after found that a version E. coli dihydrofolate reductase, which have shortened better fit into vector, is most effective. We show subretinal administration armed with short degron...
We have recently shown that dual AAV vectors expand cargo capacity in the retina up to 9kb thus allowing their application therapy of inherited blinding conditions like Stargardt disease or Usher syndrome type IB (USHIB). However, for several other size coding sequence be transferred would not fit into and require a triple vector system. generated encoding reporter EGFP-DsRed fusion protein under transcriptional control ubiquitous CMV promoter. The expression cassette was split three parts...
Abnormal accumulation of various by-products the visual cycle, including diretinoid-pyridinium-ethanolamine (A2E), in retinal pigment epithelium (RPE) is an hallmark both Stargardt disease (STGD) and age-related macular degeneration (AMD). This responsible for RPE and, consequently, photoreceptor (PR) cell death. A2E storage lysosomes has been shown to reduce capacity degrade phagocytized PR outer segments autophagosome biogenesis, trafficking autophagic flux. The transcription factor EB...