A stress-free strategy to correct point mutations in patient iPS cells.

ABSTRACT: When studying patient specific induced pluripotent stem cells (iPS cells) as a disease model, the ideal control is an isogenic line that has corrected the point mutation, instead of iPS cells from siblings or other healthy subjects. However, repairing a point mutation in iPS cells even with the newly developed CRISPR-Cas9 technique remains difficult and time-consuming. Here we report a strategy that makes the Cas9 "knock-in" methodology both hassle-free and error-free. Instead of selecting a Cas9 recognition site close to the point mutation, we chose a site located in the nearest intron. We constructed a donor template with the fragment containing the corrected point mutation as one of the homologous recombination arms flanking a PGK-Puro^R cassette. After selection with puromyci

SUBMITTER: Cai J 

PROVIDER: S-EPMC8283763 | biostudies-literature | 2021 May

REPOSITORIES: biostudies-literature