CRISPRi-Mediated Treatment of Dominant Rhodopsin-Associated Retinitis Pigmentosa

被引：6

作者：

Burnight, Erin R. ^{[1
,2
]}

Wiley, Luke A. ^{[1
,2
]}

Mullin, Nathaniel K. ^{[1
,2
]}

Adur, Malavika K. ^{[3
]}

Lang, Mallory J. ^{[1
,2
]}

Cranston, Cathryn M. ^{[1
,2
]}

Jiao, Chunhua ^{[1
,2
]}

Russell, Stephen R. ^{[1
,2
]}

Sohn, Elliot H. ^{[1
,2
]}

Han, Ian C. ^{[1
,2
]}

Ross, Jason W. ^{[3
]}

Stone, Edwin M. ^{[1
,2
]}

Mullins, Robert F. ^{[1
,2
]}

Tucker, Budd A. ^{[1
,2
]}

机构：

[1] Univ Iowa, Carver Coll Med, Inst Vis Res, Dept Ophthalmol & Visual Sci, 375 Newton Rd, Iowa City, IA 52242 USA

[2] Univ Iowa, Carver Coll Med, Dept Ophthalmol & Visual Sci, Iowa City, IA 52242 USA

[3] Iowa State Univ, Dept Anim Sci, Ames, IA USA

来源：

CRISPR JOURNAL | 2023年 / 6卷 / 06期

关键词：

LEBER CONGENITAL AMAUROSIS; CONE CELL-DEATH; GENE-THERAPY; SUBRETINAL INJECTION; PHOTORECEPTOR CELLS; RIBOZYME RESCUE; RPE65; MUTATIONS; MODEL; EXPRESSION; REPLACEMENT;

D O I：

10.1089/crispr.2023.0039

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

Rhodopsin (RHO) 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 Staphylococcus aureus Cas9 (dCas9) fused to the Kruppel-associated box (KRAB) transcriptional repressor domain. Using a reporter construct carrying green fluorescent protein (GFP) cloned downstream of the RHO promoter fragment (nucleotides -1403 to +73), we demonstrate a similar to 74-84% reduction in RHO promoter activity in RHOpCRISPRi-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 in vivo was associated with a reduction in endoplasmic reticulum (ER) stress and apoptosis markers and preservation of photoreceptor cell layer thickness.

引用

页码：502 / 513

页数：12

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