- Correction of the DMD exon 2
Correction of the exon 2 duplication in DMD myoblasts by a single CRISPR/Cas9 system. / Lattanzi, Annalisa; Duguez, Stephanie; Moiani, Arianna; Izmiryan, Araksya; Barbon, Elena; Martin, Samia; Mamchaoui, Kamel; Mouly, Vincent; Bernardi, Francesco; Mavilio, Fulvio; Bovolenta, Matteo.In: Molecular Therapy - Nucleic Acids, Vol. 7, 16.06.2017, p. 11-19.
Research output: Contribution to journal › Article
TY - JOUR
T1 - Correction of the exon 2 duplication in DMD myoblasts by a single CRISPR/Cas9 system.
AU - Lattanzi, Annalisa
AU - Duguez, Stephanie
AU - Moiani, Arianna
AU - Izmiryan, Araksya
AU - Barbon, Elena
AU - Martin, Samia
AU - Mamchaoui, Kamel
AU - Mouly, Vincent
AU - Bernardi, Francesco
AU - Mavilio, Fulvio
AU - Bovolenta, Matteo
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PY - 2017/6/16
Y1 - 2017/6/16
N2 - Exonic duplications account for 10-15% of all mutations in Duchenne muscular dystrophy (DMD), a severe hereditary neuromuscular disorder. We report a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeat)/Cas9-based strategy to correct the most frequent (exon 2) duplication in the DMD gene by targeted deletion, and tested the efficacy of such an approach in patient-derived myogenic cells. We demonstrate restoration of wild-type dystrophin expression at transcriptional and protein level in myotubes derived from genome-edited myoblasts in the absence of selection. Removal of the duplicated exon was achieved by the use of only one gRNA directed against an intronic duplicated region, thereby increasing editing efficiency and reducing the risk of off-target effects. This study opens a novel therapeutic perspective for patients carrying disease-causing duplications.
AB - Exonic duplications account for 10-15% of all mutations in Duchenne muscular dystrophy (DMD), a severe hereditary neuromuscular disorder. We report a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeat)/Cas9-based strategy to correct the most frequent (exon 2) duplication in the DMD gene by targeted deletion, and tested the efficacy of such an approach in patient-derived myogenic cells. We demonstrate restoration of wild-type dystrophin expression at transcriptional and protein level in myotubes derived from genome-edited myoblasts in the absence of selection. Removal of the duplicated exon was achieved by the use of only one gRNA directed against an intronic duplicated region, thereby increasing editing efficiency and reducing the risk of off-target effects. This study opens a novel therapeutic perspective for patients carrying disease-causing duplications.
KW - Correction of the DMD exon 2
KW - CRISPR/Cas9
U2 - 10.1016/j.omtn.2017.02.004
DO - 10.1016/j.omtn.2017.02.004
M3 - Article
VL - 7
SP - 11
EP - 19
JO - Molecular Therapy - Nucleic Acids
T2 - Molecular Therapy - Nucleic Acids
JF - Molecular Therapy - Nucleic Acids
SN - 2162-2531