Fig. 3 | Scientific Reports

Fig. 3

From: A buffer-tuning strategy to profile domain-specific activity of chimeric I-TevI/CRISPR gene editors in vitro

Fig. 3

TevCas12a cleavage of wildtype and mutated AAVS1 substrates. (A) Schematic of the linear wild-type AAVS1 substrate (SUB) indicating gRNA targeted Tev and Cas12a cleavage sites and non-targeted Tev CNNNG cleavage motifs and Cas12a TTTV PAM sequences. For simplicity, CNNNG motifs are depicted on one strand and TTTV PAM sites on the other strand. Predicted guide-targeted cleavage products are shown below. (B–E) TevCas12a cleavage timecourse assays on wild-type, \(\Delta\)CNNNG, \(\Delta\)TTTV and \(\Delta\)CNNNG/\(\Delta\)TTTV substrates. For each gel image, unreacted substrate (SUB) and targeted Tev (TP1) and Cas12a (CP1-2) cleavage products are annotated. Uncropped gel images for panels B-E are shown in Supplementary Fig. S9.

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