Premium
Genetic modification possibilities in treating corneal diseases
Author(s) -
Moore T.,
Courtney D.,
Atkinson S.,
Maurizi E.,
Allen E.,
McLean I.,
Leslie Pedrioli D.,
Moore J.,
Nesbit A.
Publication year - 2015
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2015.0145
Subject(s) - crispr , gene silencing , frameshift mutation , cas9 , transcription activator like effector nuclease , biology , mutant , genome editing , in vivo , corneal epithelium , luciferase , allele , small interfering rna , gene , gene targeting , genetics , exon , rna , transfection , epithelium
Summary Treatment of genetic eye disease poses significant medical and surgical challenges. Two gene based therapies were assessed using an in‐vivo firefly luciferase bioluminescence mouse model to target corneal epithelium. Short interfering RNA (siRNA) targeting selectively only the mutant allele was assessed for efficacy, through its ability to downregulate corneal epithelial luminescence in vivo using live animal imaging. This innovative animal model provides clear advantages enabling assessment of topical, subconjunctival and intrastromal delivery. Potent and sustained in vivo gene silencing >50% for up to 7 days was observed after intrastromal injection of siRNA and various topical formulations are further being assessed. siRNA therapy provides transient effect, however, Type II Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR‐associated nuclease 9 (Cas9) holds great promise to provide ‘one off’ permanent gene editing. Gene‐specific cleavage of the mutant allele was detected through this method. DNA repair resulted in frameshift mutations predicted to result in mutant allele knockout. This offers exciting potential for translation into clinical treatment of corneal genetic diseases.