A CRISPR–Cas9-based strategy enables the targeted introduction of homozygous or heterozygous sequence changes by homology-directed repair. The CORRECT (consecutive re-guide or re-Cas steps to erase CRISPR–Cas-blocked targets) method allows for the introduction of targeted modifications together with silent CRISPR–Cas-blocking mutations aimed at preventing 're-editing' due to high nuclease activity. Subsequent removal of the blocking mutations leaves only the intended modification. Using CORRECT, Paquet et al. generate human induced pluripotent stem cells with heterozygous and homozygous mutations in APP (which encodes amyloid-β precursor protein) and PSEN1 (which encodes presenilin 1), genes linked to the development of dominant early-onset Alzheimer disease. Cortical neurons derived from these cells exhibited disease-associated phenotypes dependent on genotype.
References
Paquet, D. et al. Efficient introduction of specific homozygous and heterozygous mutations using CRISPR/Cas9. Nature http://dx.doi.org/10.1038/nature17664 (2016)
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Koch, L. A CORRECT step forward in disease modelling. Nat Rev Genet 17, 316 (2016). https://doi.org/10.1038/nrg.2016.60
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DOI: https://doi.org/10.1038/nrg.2016.60
