saCas9 Nuclease NLS Protein

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system is the latest RNA-guided endonuclease tool in genome editing which allows for very specific genomic disruption and replacement.

The Cas9 nuclease serves to unwind the genomic DNA duplex next to conserved protospacer adjacent motifs (PAMs) and homes in on its target sequence, which is recognized by a complementary single-guide RNA. The resulting double-stranded break gets repaired by the non-homologous end joining (NHEJ) pathway, leading to a disruption in the open reading frame of the targeted gene. Alternatively, by supplying a suitable repair template, virtually any desired point mutation can be introduced at the break point via homology-directed repair (HDR).

The Cas9 nuclease from the bacteria Staphylococcus aureus, abbreviated saCas9, is gaining popularity as an alternative to spCas9 due to its relatively smaller size. The saCas9 PAM sequence is 5’-NNGRRN (preferably 5’-NNGRRT). saCas9 Nuclease NLS contains a SV40 T antigen nuclear localization sequence (NLS) on the C-terminus of the protein.