Extended Data Figure 5. dCas9^St can also support spacer acquisition.

A plasmid derived from pRH241 containing mutations in the active site of St Cas9 (D10A, H847A; dCas9^St) was used to characterize spacer acquisition in the absence of phage infection. Upon over-expression of Cas1, Cas2 and Csn2 using anydrotetracycline (aTc), we were able to detect spacer acquisition. Sequencing of spacers and alignment of the protospacer flanking sequences demonstrated the selection of an NGGNG PAM. Image is representative of three technical replicates.