Cas9 nuclease (4 μg/μl) for the generation of double-strand breaks in CRISPR-Cas9 genome editing.

ArciTect™ Cas9 Nuclease requires association with a guide RNA—e.g. ArciTect™ sgRNA (Catalog #STMC200-0013) or a duplex composed of ArciTect™ tracrRNA (Catalog #STMC76016) and ArciTect™ crRNA (Catalog #STMC76010)—to form a ribonucleoprotein (RNP) complex. This RNP complex then creates double-strand breaks at site-specific locations in the genome. ArciTect™ Cas9 Nuclease contains a nuclear localisation signal at the N-terminus, ensuring that the RNP complex translocates to the nucleus, thereby increasing the efficiency of genome editing. As the RNP complex is fully functional upon transfection, it allows for immediate activity following translocation to the nucleus. The RNP complex is degraded over 48 hours, allowing sufficient time for genome editing to occur while reducing off-target effects that can be caused by the continuous presence of the RNP complex. Using the RNP system also circumvents the laborious process of generating stable Cas9-expressing cell lines, saving time and reducing the risk of off-target effects due to leaky inducible expression systems. The S. pyogenes Cas9 uses the protospacer adjacent motif (PAM) sequence NGG (where N can be any nucleotide). The enzyme will not cleave without a genomic PAM site downstream of the target sequence.