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. 2026 Apr 13;33(5):756–767. doi: 10.1038/s41594-026-01788-6

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© The Author(s) 2026

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a, Schematic diagram of gRNA scaffolds of OsCas12f, RhCas12f and Al3Cas12f. The tracrRNA, crRNA and tetraloop are labeled and colored accordingly. Unresolved regions are indicated by a dashed line. b, Surface views of OsCas12f, RhCas12f and Al3Cas12f bound to gRNA. Dotted circles highlight the differences of stem 2 in OsCas12f and RhCas12f gRNA. This region is present in RhCas12f gRNA because of the PAM-analogous contacts highlighted in c. c, Zoomed-in overview of interactions between the RhCas12f WED/REC domains and stem 2 of its cognate gRNA. d, Plasmid interference activity of wild-type and gRNA truncation mutants in OsCas12f and RhCas12f. Data represent the mean ± s.d. (n = 3 independent biological replicates). Significance was determined using a one-way analysis of variance (ANOVA) for OsCas12f or unpaired t-test for RhCas12f.

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