Rapid repair of human disease-specific single-nucleotide variants by One-SHOT genome editing
Many human diseases ranging from cancer to hereditary disorders are caused by single-nucleotide mutations in critical genes. Repairing these mutations would significantly improve the quality of life for patients with hereditary diseases. However, current procedures for repairing deleterious single-n...
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Veröffentlicht in: | Scientific reports 2020-08, Vol.10 (1), p.13927-13927, Article 13927 |
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Zusammenfassung: | Many human diseases ranging from cancer to hereditary disorders are caused by single-nucleotide mutations in critical genes. Repairing these mutations would significantly improve the quality of life for patients with hereditary diseases. However, current procedures for repairing deleterious single-nucleotide mutations are not straightforward, requiring multiple steps and taking several months to complete. In the current study, we aimed to repair pathogenic allele-specific single-nucleotide mutations using a single round of genome editing. Using high-fidelity, site-specific nuclease
As
Cas12a/Cpf1, we attempted to repair pathogenic single-nucleotide variants (SNVs) in disease-specific induced pluripotent stem cells. As a result, we achieved repair of the Met918Thr SNV in human oncogene
RET
with the inclusion of a single-nucleotide marker, followed by absolute markerless, scarless repair of the
RET
SNV with no detected off-target effects. The markerless method was then confirmed in human type VII collagen-encoding gene
COL7A1
. Thus, using this One-SHOT method, we successfully reduced the number of genetic manipulations required for genome repair from two consecutive events to one, resulting in allele-specific repair that can be completed within 3 weeks, with or without a single-nucleotide marker. Our findings suggest that One-SHOT can be used to repair other types of mutations, with potential beyond human medicine. |
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ISSN: | 2045-2322 2045-2322 |
DOI: | 10.1038/s41598-020-70401-7 |