The first step in your experimental design is to select an appropriate CRISPR-Cas system. The most commonly used variant is the CRISPR-Cas9 system and if you haven't performed any genome editing experiments yet, this is the best starting point. Depending on your experimental goals and the genes you are trying to target, it may me worthwhile to also try other CRISPR systems, such as the CRISPR-Cas12a system.
The CRISPR-Cas9 system
This is the current gold standard — start with this system if this is your first attempt at editing your gene of interest.
Required PAM site: NGG
Benefit from the latest improvements in on- and off-target design and chemical modifications
Get optimal editing with high on-target potency and reduced off-target activity
Precisely control editing with efficient delivery of the RNP by lipofection or electroporation
The CRISPR-Cas12a system
The Alt-R CRISPR-Cas12a System allows targeting of alternative sites that are not available to the CRISPR-Cas9 system and produces a staggered cut with a 5’ overhang. May work better than CRISPR-Cas9 for AT-rich genomes.
Required PAM site: TTTV (V = A, C or G)
Enables genome editing in organisms with AT-rich genomes
Allows interrogation of additional genomic regions compared to Cas9
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