SpCas9 nickase vectors are advantageous to use in experiments which are more sensitive to off-target editing. However, it is important to remember, that two gRNAs will need to be designed to target both forward and reverse strands. These gRNAs must be oriented so that PAM sites are distal to each other. gRNA sequences should be offset with windows of up to 100bp between them.
Articles in this section
- What are the factors that can affect CRISPR targeting efficiency and specificity?
- What expression vectors should I use?
- What are your CRISPR plasmid delivery specifications?
- How many gRNA sequences are needed for targeted knock-out?
- How should gRNA sequences be designed?
- Do I need an all-in-one or dual vector system?
- When are lentiviral or adeno-associated viral (AAV) vectors necessary?
- When should I use SpCas9 nickase vectors?
- How are gRNA sequences designed for the transcription activation (SAM) system?