FIG 4 .

Orphan CRISPR2 provides defense against PRPs in the presence of CRISPR1 cas9. (A) Top panel: schematic of how the T11 CRISPR2 spacer 1 sequence and corresponding PAM sequences (underlined) were introduced into pCF10. (Bottom panel) Conjugation frequencies of T11RF and its derivatives as recipient strains in conjugation with OG1SSp harboring pCF10, pVP501, and pVP502. The T11 CRISPR2 locus provides genome defense against pVP501 in the presence of CRISPR1 cas9 and its predicted tracrRNA. Results also demonstrate CRISPR1 cas9 PAM specificity to the NGG sequence. A minimum of three independent conjugation reactions are represented. Significance was assessed using a one-tailed Student’s t test; P values are relative to pVP501 transfer to Δcas9+CR1: **, P < 0.005. (B) Outline of donor and recipient strains used for assessing the function of CRISPR2.