Fig. 4: NUC lobe of apoCas9 is sufficient for super-adaptation but requires the REC lobe for RNA regulation.
From: Cas9 senses CRISPR RNA abundance to regulate CRISPR spacer acquisition
a, Schematics of NmeCas9 domain architecture and serial deletion mutants. aa, amino acid. b, NUC lobe of NmeCas9, which lacks bridge helix (BH) and REC1/2 domains, is sufficient for super-adaptation. Top, a representative adaptation PCR gel. Bottom, quantification of adaptation efficiencies. Data are mean ± s.d., n = 3. NS (P ≥ 0.05), *0.005 ≤ P < 0.05 and **P < 0.005; P values calculated using two-tailed Welch’s t-tests. c, 3′ Flanking motif analysis for new viral spacers of b. d,NmeCas9 mutants ΔREC1/2 and ΔBHΔREC1/2 became resistant to RNA regulation. Top, a representative adaptation PCR gel. Bottom, quantification of adaptation efficiencies. Data are mean ± s.d., n = 3. NS (P ≥ 0.05), *0.005 ≤ P < 0.05 and **P < 0.005. e, 3′ Flanking motif analysis for new viral spacers of d.
