Journal article
CRISPRi-Mediated Treatment of Dominant Rhodopsin-Associated Retinitis Pigmentosa
CRISPR journal, Vol.6(6), pp.502-513
12/2023
DOI: 10.1089/crispr.2023.0039
PMCID: PMC11304754
PMID: 38108516
Abstract
Rhodopsin (
) mutations such as Pro23His are the leading cause of dominantly inherited retinitis pigmentosa in North America. As with other dominant retinal dystrophies, these mutations lead to production of a toxic protein product, and treatment will require knockdown of the mutant allele. The purpose of this study was to develop a CRISPR-Cas9-mediated transcriptional repression strategy using catalytically inactive
Cas9 (dCas9) fused to the Krüppel-associated box (KRAB) transcriptional repressor domain. Using a reporter construct carrying green fluorescent protein (GFP) cloned downstream of the
promoter fragment (nucleotides -1403 to +73), we demonstrate a ∼74-84% reduction in
promoter activity in
CRISPRi-treated versus plasmid-only controls. After subretinal transduction of human retinal explants and transgenic Pro23His mutant pigs, significant knockdown of rhodopsin protein was achieved. Suppression of mutant transgene
was associated with a reduction in endoplasmic reticulum (ER) stress and apoptosis markers and preservation of photoreceptor cell layer thickness.
Details
- Title: Subtitle
- CRISPRi-Mediated Treatment of Dominant Rhodopsin-Associated Retinitis Pigmentosa
- Creators
- Erin R Burnight - University of IowaLuke A Wiley - University of IowaNathaniel K Mullin - University of IowaMalavika K Adur - Iowa State UniversityMallory J Lang - University of IowaCathryn M Cranston - University of IowaChunhua Jiao - Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, Iowa, USAStephen R Russell - University of IowaElliot H Sohn - Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, Iowa, USAIan C Han - University of IowaJason W Ross - Iowa State UniversityEdwin M Stone - University of IowaRobert F Mullins - University of IowaBudd A Tucker - Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, Iowa, USA
- Resource Type
- Journal article
- Publication Details
- CRISPR journal, Vol.6(6), pp.502-513
- DOI
- 10.1089/crispr.2023.0039
- PMID
- 38108516
- PMCID
- PMC11304754
- NLM abbreviation
- CRISPR J
- eISSN
- 2573-1602
- Language
- English
- Date published
- 12/2023
- Academic Unit
- The University of Iowa Institute for Vision Research; Iowa Neuroscience Institute; John and Marcia Carver Nonprofit Genetic Testing Laboratory; Fraternal Order of Eagles Diabetes Research Center; Ophthalmology and Visual Sciences
- Record Identifier
- 9984533289102771
Metrics
22 Record Views