Spcas9 has been reported to cleave ~ 3bp upstream of the start of the PAM as a blunt cut, Cpf1 has a staggered cut significantly upstream of the PAM, but I cant seem to find much information regarding the cleavage site of SaCas9, anyone know of any potentially helpful publications?
Hi,
like SpCas9, most Cas9 orthologues including SaCas9 were shown to cut three base pairs upstream of their respective PAM sequence (in case of SaCas9 5'-NNGRRT-3'). For more detailed information you can look here:
- Ran, F., Cong, L., Yan, W. et al. In vivo genome editing using Staphylococcus aureus Cas9. Nature 520, 186–191 (2015). https://doi.org/10.1038/nature14299
- Steinert J., Schiml S., Fauser F. and Puchta H. (2015) Highly efficient heritable plant genome engineering using Cas9 orthologues from Streptococcus thermophilus and Staphylococcus aureus. Plant J. DOI: 10.1111/tpj.13078
Seems to be more frequent at 3-bp upstream of PAM, according to this paper:
"In vivo genome editing using Staphylococcus aureus Cas9", https://doi.org/10.1038/nature14299
Extended figure 2
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