Cystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the CFTR gene. The 3272–26A>G and 3849+10kbC>T CFTR mutations alter the correct splicing of the CFTR gene, generating new acceptor and donor splice sites respectively. Here we develop a genome editing approach to permanently correct these genetic defects, using a single crRNA and the Acidaminococcus sp. BV3L6, AsCas12a. This genetic repair strategy is highly precise, showing very strong discrimination between the wild-type and mutant sequence and a complete absence of detectable off-targets. The efficacy of this gene correction strategy is verified in intestinal organoids and airway epithelial cells derived from CF patients carrying the 3272–26A>G or 3849+10kbC>T mutations, showing efficient repair and complete functional recovery of the CFTR channel. These results demonstrate that allele-specific genome editing with AsCas12a can correct aberrant CFTR splicing mutations, paving the way for a permanent splicing correction in genetic diseases.

Allele specific repair of splicing mutations in cystic fibrosis through AsCas12a genome editing / Maule, G.; Casini, A.; Montagna, C.; Ramalho, A. S.; De Boeck, K.; Debyser, Z.; Carlon, M. S.; Petris, G.; Cereseto, A.. - In: NATURE COMMUNICATIONS. - ISSN 2041-1723. - 10:1(2019), pp. 355601-355611. [10.1038/s41467-019-11454-9]

Allele specific repair of splicing mutations in cystic fibrosis through AsCas12a genome editing

Maule G.;Casini A.;Montagna C.;Petris G.;Cereseto A.
2019-01-01

Abstract

Cystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the CFTR gene. The 3272–26A>G and 3849+10kbC>T CFTR mutations alter the correct splicing of the CFTR gene, generating new acceptor and donor splice sites respectively. Here we develop a genome editing approach to permanently correct these genetic defects, using a single crRNA and the Acidaminococcus sp. BV3L6, AsCas12a. This genetic repair strategy is highly precise, showing very strong discrimination between the wild-type and mutant sequence and a complete absence of detectable off-targets. The efficacy of this gene correction strategy is verified in intestinal organoids and airway epithelial cells derived from CF patients carrying the 3272–26A>G or 3849+10kbC>T mutations, showing efficient repair and complete functional recovery of the CFTR channel. These results demonstrate that allele-specific genome editing with AsCas12a can correct aberrant CFTR splicing mutations, paving the way for a permanent splicing correction in genetic diseases.
2019
1
Maule, G.; Casini, A.; Montagna, C.; Ramalho, A. S.; De Boeck, K.; Debyser, Z.; Carlon, M. S.; Petris, G.; Cereseto, A.
Allele specific repair of splicing mutations in cystic fibrosis through AsCas12a genome editing / Maule, G.; Casini, A.; Montagna, C.; Ramalho, A. S.; De Boeck, K.; Debyser, Z.; Carlon, M. S.; Petris, G.; Cereseto, A.. - In: NATURE COMMUNICATIONS. - ISSN 2041-1723. - 10:1(2019), pp. 355601-355611. [10.1038/s41467-019-11454-9]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11572/287753
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