Dear All, I am now designing the sgRNA for knocking out the gene through crispr. Base on my learning experience, a start codon and a stop codon should be put in the N terminus and the C terminus of the DNA sequence for transcription. However, for sgRNA, many of the website just said adding the crRNA to the tracRNA in the presence of a promoter is fine, but I dont know if the start codon and stop codon are needed to be added for the sgRNA transcription. Hope you can help me to answer the question. Thank you.