High-Efficiency CRISPR/Cas9-Mediated Correction of a Homozygous Mutation in Achromatopsia-Patient-Derived iPSCs

Abstract

Achromatopsia is an autosomal recessive disorder, in which cone photoreceptors undergo progressive degeneration, causing color blindness and poor visual acuity, among other significant eye affectations. It belongs to a group of inherited retinal dystrophies thatcurrently have no treatment. Although functional improvements have been reported in several ongoing gene therapy studies, more efforts and research should be carried out to enhance their clinical application. In recent years, genome editing has arisen as one of the most promising tools for personalized medicine. In this study, we aimed to correct a homozygous PDE6C pathogenic variant in hiPSCs derived from a patient affected by achromatopsia through CRISPR/Cas9 and TALENs technologies. Here, we demonstrate high efficiency in gene editing by CRISPR/Cas9 but not with TALENs approximation. Despite a few of the edited clones displaying heterozygous on-target defects, the proportion of corrected clones with a potentially restored wild-type PDE6C protein was more than half of the total clones analyzed. In addition, none of them presented off-target aberrations. These results significantly contribute to advances in single-nucleotide gene editing and the development of future strategies for the treatment of achromatopsia.

Endoplasmic reticulum stress: molecular mechanism and therapeutic targets

“…This can be avoided by using CRISPR/Cas9-mediated gene editing to correct single-nucleotide mutations for PDE6. 357 ER STRESS AND CATARACTS Cataracts refer to lens opacification and remain the leading cause of blindness in middle-and low-income countries, 358 affecting approximately 94 million people worldwide. In 2020, about 15 million people over the age of 50 worldwide will be blind due to cataracts, and about 79 million will have moderate-to-severe visual impairment.…”

CRISPR/Cas9: implication for modeling and therapy of amyotrophic lateral sclerosis

“…TALENs, cheaper, safer, more efficient, and easier to engineer than zinc-finger proteins, can be applied more widely to life sciences than ZFNs (Huo et al, 2019). This gene-editing tool was utilized for various cell lines, including induced pluripotent stem cells (iPSCs) (Kwon et al, 2021;Qiu et al, 2021;Siles et al, 2023), nematodes (Verdu et al, 2020(Verdu et al, , 2021, zebrafish (Zhu et al, 2021;Sertori et al, 2022), and generating knockouts in rodent models (Lee et al, 2021). Despite the improvement, simplification, and high specificity of TALENs, they have an obvious limitation: being quite large (impeding delivery).…”