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Genome Editing in Patient iPSCs Corrects the Most Prevalent USH2A Mutations and Reveals Intriguing Mutant mRNA
Carla Sanjurjo-Soriano1,2, Nejla Erkilic1,2, David Baux2,3
1Inserm U1051, Institute for Neurosciences of Montpellier, 34091 Montpellier, France.
Molecular Therapy. Methods & Clinical Development
|January 8, 2020
Summary
CRISPR/Cas9 genome editing successfully corrected common USH2A mutations in patient stem cells, offering a potential treatment for Usher syndrome (USH) and autosomal recessive retinitis pigmentosa (arRP). This approach shows high efficacy and specificity without off-target effects.
Area of Science:
- Genetics
- Molecular Biology
- Ophthalmology
Background:
- Inherited retinal dystrophies (IRDs) cause progressive vision loss.
- Usher syndrome (USH) involves retinitis pigmentosa (RP) and hearing loss, often caused by USH2A gene mutations.
- Two prevalent USH2A mutations (c.2276G>T and c.2299delG) are responsible for many USH and isolated autosomal recessive RP (arRP) cases.
Purpose of the Study:
- To investigate the feasibility of CRISPR/Cas9 genome editing for correcting prevalent USH2A mutations.
- To assess the efficacy, specificity, and safety of enhanced specificity Cas9 (eSpCas9) in patient-derived cells.
Main Methods:
- Utilized eSpCas9 to precisely correct c.2276G>T and c.2299delG USH2A mutations in induced pluripotent stem cells (iPSCs).
- Evaluated target on-target efficacy and screened for off-target mutagenesis.
- Assessed pluripotency and genetic stability of corrected iPSCs.
- Analyzed USH2A mRNA expression levels before and after correction.
Main Results:
- Achieved seamless correction of the two recurrent USH2A mutations in patient iPSCs.
- Demonstrated high target efficacy and specificity of eSpCas9 with no detectable off-target mutations.
- Confirmed retention of pluripotency and genetic stability in corrected iPSCs.
- Observed reversion of aberrant USH2A mRNA levels following mutation correction.
Conclusions:
- CRISPR/Cas9-mediated genome editing is a viable and efficient strategy for correcting prevalent USH2A mutations.
- This approach holds promise for developing potential therapeutic interventions for Usher syndrome and autosomal recessive RP.
- The study highlights the potential of eSpCas9 for precise gene correction in inherited retinal diseases.
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