Extended Data Fig. 3: Additional analysis of dCas12f-associated gRNAs from RIP–seq experiments and comparative genomics analyses.

a, RIP–seq coverage plots for the five indicated dCas12f homologues, revealing a well-defined gRNA with scaffold (orange) and guide (purple) regions. Nucleotides outside the boundary of the presumed full-length gRNA are coloured in grey. Total gRNA lengths are noted to the right of each plot. The guide region of Zunongwangia profunda is plotted on a separate y-axis scale, for visual clarity. Coverage is shown as counts per million (CPM). b, Zoomed-out RIP–seq coverage plots for an additional dCas12f orthologue from Leeuwenhoekiella palythoae (Lpa), showing both a zoomed-out view (left), depicting the same data as shown in Fig. 2c, and magnified view as in a (right). Region 1 annotated in the operon schematic (top left) encodes multiple tandem gRNAs with similar scaffold and guide sequences. Coverage is shown as in a. c, RIP–seq coverage plots for an additional dCas12f orthologue from Paenimyroides ummariense (Pum), shown as in b, with the magnified view at right comparing the aligned scaffold and guide sequences. The first two gRNA sequences are identical, and bases that differ in the third gRNA are highlighted in red. d, Annotated genomic neighborhood of an rpoE-dcas12f operon that encodes both predicted full-length gRNAs (orange/purple) and multiple discontinuous CRISPR arrays (repeats in tan). The sequence similarity between the CRISPR repeats and gRNA scaffold (grey circles, bottom) suggests a potential evolutionary emergence of chimeric, dCas12f-associated single guide RNAs from CRISPR arrays.