Fiche publication
Date publication
septembre 2020
Journal
Stem cell reports
Auteurs
Membres identifiés du Cancéropôle Est :
Dr CULLOT Grégoire
Tous les auteurs :
Prat F, Toutain J, Boutin J, Amintas S, Cullot G, Lalanne M, Lamrissi-Garcia I, Moranvillier I, Richard E, Blouin JM, Dabernat S, Moreau-Gaudry F, Bedel A
Lien Pubmed
Résumé
CRISPR/Cas9 is a promising technology for gene correction. However, the edition is often biallelic, and uncontrolled small insertions and deletions (indels) concomitant to precise correction are created. Mutation-specific guide RNAs were recently tested to correct dominant inherited diseases, sparing the wild-type allele. We tested an original approach to correct compound heterozygous recessive mutations. We compared editing efficiency and genotoxicity by biallelic guide RNA versus mutant allele-specific guide RNA in iPSCs derived from a congenital erythropoietic porphyria patient carrying compound heterozygous mutations resulting in UROS gene invalidation. We obtained UROS function rescue and metabolic correction with both guides with the potential of use for porphyria clinical intervention. However, unlike the biallelic one, the mutant allele-specific guide was free of on-target collateral damage. We recommend this design to avoid genotoxicity and to obtain on-target scarless gene correction for recessive disease with frequent cases of compound heterozygous mutations.
Mots clés
CRISPR/Cas9, compound heterozygous recessive disease, congenital erythropoietic porphyria, gene therapy, iPSCs, mutation specific, precise genome editing
Référence
Stem Cell Reports. 2020 09 8;15(3):677-693
