Authors
Ajay Gupta, Rabia Ahuja, Bo Liu, Mark Adero, Dung Thi Pham, Wolf B Frommer, Bing Yang
Published in
aBIOTECH. Volume 7. Issue 3. Pages 100054. Epub May 20, 2026.
Abstract
Recent advances in generative artificial intelligence (AI) have enabled the de novo design of genome-editing nucleases. For example, OpenCRISPR-1 offers an open-source alternative to naturally evolved CRISPR systems and expands the "freedom to operate" (FTO). Here, we report the development and systematic validation of a monocot-optimized OpenCRISPR-1-based genome-editing ecosystem in rice (Oryza sativa). By targeting the OsSWEET susceptibility (S) gene family, we demonstrate that OpenCRISPR-1 supports robust multiplexed editing in both rice calli and stable T0 plants, with mutation frequencies reaching 100% in some samples. Deep sequencing revealed that the OpenCRISPR-1 mutational landscape mirrors that of Streptococcus pyogenes Cas9 (SpCas9), facilitating the development of predictable loss-of-function alleles that confer broad-spectrum resistance to bacterial blight. To develop a fully open-source platform, we integrated an AI-designed Open sgRNA scaffold (OpsgRNA), which maintained high editing efficacy across multiple target loci, into our editing system. Furthermore, we expanded the toolkit by engineering OpenPE6c, an OpenCRISPR-1-based prime editing system. OpenPE6c exhibited precise editing rates in rice protoplasts comparable to that of canonical SpCas9-PE6c while significantly reducing imprecise byproducts, suggesting that the AI-designed nuclease has enhanced fidelity. Our results establish OpenCRISPR-1 as a versatile, high-performance, public-access platform for advanced plant genome engineering, offering a transparent framework for the global democratization of precision crop breeding.
PMID:
42434514
Bibliographic data and abstract were imported from PubMed on 11 Jul 2026.
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