Scarless introduction of point mutations in mammalian cells with S. pyogenes Cas9 v1

This protocol outlines the steps to edit single point mutations into cells in a scarless manner. Uses single-stranded DNA repair templates and the S. pyogenes CRISPR/Cas9 system. Using a short single-stranded DNA repair template is advantageous because it is easy to order synthetically and it cannot be randomly integrated into other double-strand breaks in the cell. Furthermore, this protocol is "scarless" - that is, nothing is changed to the genome other than the desired changes. However, it may be easier to edit genes using a selectable marker, especially when knocking out a gene. This protocol is ideal when trying to make genetic changes with minimal perturbations to the gene locus and surroundings. Note that repair using a single-stranded short repair template (SSTR) uses a different DNA repair pathway than homology-directed repair from long DNA repair templates. Citations: image 1: synthego.com- verify your guide design tool