RNA-Guided Genome Editing of Mammalian Cells

The microbial CRISPR-Cas adaptive immune system can be harnessed to facilitate genome editing in eukaryotic cells (Cong L et al., Science 339, 819–823, 2013; Mali P et al., Science 339, 823–826, 2013). Here we describe a protocol for the use of the RNA-guided Cas9 nuclease from the Streptococcus pyogenes type II CRISPR system to achieve specifi c, scalable, and cost-effi cient genome editing in mammalian cells. Key words CRISPR-Cas Genome editing DNA cleavage Cas9 Guide RNA PAM sequence NHE