CRISPR/Cas9-based precise excision of SlHyPRP1 domain(s) to obtain salt stress-tolerant tomato
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ORIGINAL ARTICLE
CRISPR/Cas9‑based precise excision of SlHyPRP1 domain(s) to obtain salt stress‑tolerant tomato Mil Thi Tran1,2 · Duong Thi Hai Doan1 · Jihae Kim1 · Young Jong Song1 · Yeon Woo Sung1 · Swati Das1 · Eun‐ Jung Kim1,3 · Geon Hui Son1 · Sang Hee Kim1 · Tien Van Vu1,4 · Jae‑Yean Kim1,5 Received: 29 June 2020 / Accepted: 3 October 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Key message CRISPR/Cas9-based multiplexed editing of SlHyPRP1 resulted in precise deletions of its functional motif(s), thereby resulting in salt stress-tolerant events in cultivated tomato. Abstract Crop genetic improvement to address environmental stresses for sustainable food production has been in high demand, especially given the current situation of global climate changes and reduction of the global food production rate/population rate. Recently, the emerging clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas)-based targeted mutagenesis has provided a revolutionary approach to crop improvement. The major application of CRISPR/Cas in plant genome editing has been the generation of indel mutations via error-prone nonhomologous end joining (NHEJ) repair of DNA DSBs. In this study, we examined the power of the CRISPR/Cas9-based novel approach in the precise manipulation of protein domains of tomato hybrid proline-rich protein 1 (HyPRP1), which is a negative regulator of salt stress responses. We revealed that the precise elimination of SlHyPRP1 negative-response domain(s) led to high salinity tolerance at the germination and vegetative stages in our experimental conditions. CRISPR/Cas9-based domain editing may be an efficient tool to engineer multidomain proteins of important food crops to cope with global climate changes for sustainable agriculture and future food security. Keywords CRISPR/Cas9 · Genome editing · HyPRP1 · Multiplexed editing · Precision breeding · Tomato
Introduction Communicated by Günther Hahne. Mil Thi Tran, Duong Thi Hai Doan and Jihae Kim have equally contributed to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00299-020-02622-z) contains supplementary material, which is available to authorized users. * Tien Van Vu [email protected] * Jae‑Yean Kim [email protected] 1
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Division of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju 660‑701, Republic of Korea
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) cleavage usually produces blunt-end double-stranded breaks (DSBs) at a position three base pairs upstream of the NGG PAM sequence (Jinek et al. 2012; Van Vu et al. 2019). The DSB blunt ends are efficiently religated via nonhomologous 3
Present Address: Department of Life Sciences, Pohang University of Science and Technology, Pohang, Gyeongbuk, Republic of Korea
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National Key Laboratory for Plant Cell Bio
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