AAV Gene Augmentation Therapy for CRB1-Associated Retinitis Pigmentosa

C Henrique Alves; J. Wijnholds

doi:10.1007/978-1-4939-7522-8_10

AAV Gene Augmentation Therapy for CRB1-Associated Retinitis Pigmentosa

NIN

Onderzoeksoutput: Bijdrage aan wetenschappelijk tijdschrift/periodieke uitgave › Artikel › Wetenschappelijk › peer review

Samenvatting

Mutations in the CRB1 gene account for around 10,000 persons with Leber congenital amaurosis (LCA) and 70,000 persons with retinitis pigmentosa (RP) worldwide. Therefore, the CRB1 gene is a key target in the fight against blindness. A proof-of-concept for an adeno-associated virus (AAV)-mediated CRB2 gene augmentation therapy for CRB1-RP was recently described. Preclinical studies using animal models such as knockout or mutant mice are crucial to obtain such proof-of-concept. In this chapter we describe a technique to deliver AAV vectors, into the murine retinas, via the subretinal route. We also present protocols to detect expression of the therapeutic protein by fluorescence immunohistochemistry and to perform histological studies using ultra-thin sections stained with toluidine blue. These techniques in combination with electroretinography and visual behavior tests are in principle sufficient to obtain proof-of-concept for new gene therapies.

Originele taal-2	Engels
Pagina's (van-tot)	135-151
Aantal pagina's	17
Tijdschrift	Methods in Molecular Biology
Volume	1715
DOI's	https://doi.org/10.1007/978-1-4939-7522-8_10
Status	Gepubliceerd - 2018

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10.1007/978-1-4939-7522-8_10

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Citeer dit

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