P.F508del editing in cells from cystic fibrosis patients.

Autor: Smirnikhina SA; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russian Federation., Kondrateva EV; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russian Federation., Adilgereeva EP; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russian Federation., Anuchina AA; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russian Federation., Zaynitdinova MI; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russian Federation., Slesarenko YS; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russian Federation., Ershova AS; Nazarbayev University, School of Science and Technology, Nur-Sultan, Republic of Kazakhstan., Ustinov KD; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russian Federation., Yasinovsky MI; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russian Federation., Amelina EL; Laboratory of Cystic Fibrosis, Research Institute of Pulmonology, Moscow, Russian Federation., Voronina ES; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russian Federation., Yakushina VD; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russian Federation., Tabakov VY; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russian Federation., Lavrov AV; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russian Federation.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2020 Nov 11; Vol. 15 (11), pp. e0242094. Date of Electronic Publication: 2020 Nov 11 (Print Publication: 2020).
DOI: 10.1371/journal.pone.0242094
Abstrakt: Development of genome editing methods created new opportunities for the development of etiology-based therapies of hereditary diseases. Here, we demonstrate that CRISPR/Cas9 can correct p.F508del mutation in the CFTR gene in the CFTE29o- cells and induced pluripotent stem cells (iPSCs) derived from patients with cystic fibrosis (CF). We used several combinations of Cas9, sgRNA and ssODN and measured editing efficiency in the endogenous CFTR gene and in the co-transfected plasmid containing the CFTR locus with the p.F508del mutation. The non-homologous end joining (NHEJ) frequency in the CFTR gene in the CFTE29o- cells varied from 1.25% to 2.54% of alleles. The best homology-directed repair (HDR) frequency in the endogenous CFTR locus was 1.42% of alleles. In iPSCs, the NHEJ frequency in the CFTR gene varied from 5.5% to 12.13% of alleles. The best HDR efficacy was 2.38% of alleles. Our results show that p.F508del mutation editing using CRISPR/Cas9 in CF patient-derived iPSCs is a relatively rare event and subsequent cell selection and cultivation should be carried out.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
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