Cas9 Nuclease GFP 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 from the bacteria Streptococcus pyogenes, abbreviated spCas9, is the most commonly used Cas9 variant. The fusion of Cas9 Nuclease NLS to GFP allows for visual confirmation of transfection as well as subsequent verification of Cas9 clearance from the cells. Cas9 Nuclease-GFP can also be used for FACS applications and screening. Cas9 Nuclease-GFP NLS contains a SV40 T antigen nuclear localization sequence (NLS) on the C-terminus of the protein. Our Cas9-GFP proteins have the Enhanced green fluorescent protein (eGFP).