The single guide RNA for use with EnGen Seq1 Cas9 has the following sequence:
5’- NNNNNNNNNNNNNNNNNNNNGUUUUAGAGCUGUGUUGGAAACAACACAGCGAGUUAAAAUAAGGCUUUGUCCGUACACAACUUGUAAAAGUGGCACCCGAUUCGGGUGCAUUUUUU -3’
The underlined 20-nucleotide spacer sequence is specific to the desired DNA target region immediately upstream of the 5’-NAGA-3’ protospacer adjacent motif. The spacer sequence does not contain the PAM sequence itself. The portion of the sgRNA not underlined is the scaffold specific for ribonucleoprotein complex formation with EnGen Seq1 Cas9.
