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BRIEF RESEARCH REPORT article

Front. Genome Ed.
Sec. Genome Editing in Plants
Volume 6 - 2024 | doi: 10.3389/fgeed.2024.1506468
This article is part of the Research Topic Developments in Genetic Transformation and Genome Editing in Horticultural Crops and Their Pathogens View all articles

Intein-mediated split SaCas9 for genome editing in plants

Provisionally accepted
Danling Hu Danling Hu Lizhe Hu Lizhe Hu Yaqiang Lu Yaqiang Lu Xiao Dong Xiao Dong Xingyu Cao Xingyu Cao Shasha Bai Shasha Bai Lingang Zhang Lingang Zhang Dongming Li Dongming Li Yongwei Sun Yongwei Sun *
  • Inner Mongolia University, Hohhot, China

The final, formatted version of the article will be published soon.

    Virus-induced genome editing (VIGE) technologies have been developed to address the limitations of plant genome editing, which heavily relies on genetic transformation and regeneration. However, the application of VIGE in plants is hampered by the challenge posed by the size of the commonly used gene-editing nucleases, Cas9 and Cas12a. To overcome this challenge, we employed intein-mediated protein splicing to divide the SaCas9 transcript into two segments (Split-v1) and three segments (Split-v3). The Split-v1 system demonstrated genome editing efficiencies in transgenic plants comparable to those achieved with wild-type SaCas9, with efficiencies ranging from 70.2% to 96.1%. Additionally, we constructed barley stripe mosaic virus (BSMV)-based vectors to co-express Split-v1 SaCas9 and gRNAs targeting LcHRC, LcGW2, and LcTB1 in sheepgrass(Leymus chinensis), a Gramineae forage species known for its recalcitrance to genetic transformation.Infected leaves of sheepgrass exhibited genome editing efficiencies ranging from 10.40% to 37.03%. These results demonstrate the potential of intein-mediated split nuclease systems to broaden the applicability of VIGE in challenging plant species.

    Keywords: CRiSPR/Cas, intein, SaCas9, VIGE, Plants

    Received: 05 Oct 2024; Accepted: 13 Dec 2024.

    Copyright: © 2024 Hu, Hu, Lu, Dong, Cao, Bai, Zhang, Li and Sun. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Yongwei Sun, Inner Mongolia University, Hohhot, China

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