The binary vectors contain Cas9 (nuclease or nickase) and the gRNA. Cas9 is codon optimized for high expression in monocots and dicots.
Cas9 nuclease vector for single and multiple target sequences
RNA is driven by the U6 promoter from Arabidopsis thaliana (for dicots) or Oryza sativa (for monocots).
Cas9 nickase vector
Insertion of the target RNA in the Cas9 nickase vector can be easily done as DNA fragment containing both target RNAs at the ends. The inserted DNA fragment contains the U6 promoter.
|Cloning of two target sites into binary nickase or double nuclease vector|