Abstract
Key message
4382 available sgRNAs targeting 1060 tobacco genes were obtained, and 10,682 targeted mutants were created using high-throughput methods. Four optimization experiments were established to solve problems encountered during genetic transformation.
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Data from this research that were also reviewed are included in the supplementary materials.
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Funding
This work was supported by [Creation and Identification of Editing Material Library of Tobacco Transporter and Enzyme-Related Genes in Red flower Mammoth Gold] (Grant number [110202101034] to H.X.) and [Development of Tagless Gene Editing Technology and Breeding Application of Low Nicotine Editing Materials] (Grant number [2022JY03] to H.X.).
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YC: Conceptualization, visualization, writing- original draft preparation, writing—review and editing. HX: Conceptualization, validation, funding acquisition, writing—original draft preparation, writing—review and editing. LJ: Investigation, writing—review and editing. QY: Writing—review and editing. JZ: Visualization, writing—review and editing. JJ: Investigation, writing—review and editing. WZ: Supervision, writing—review and editing. LD: Investigation. JJ: Investigation, writing—review and editing. QG: Conceptualization, validation, writing—review and editing. XL: Conceptualization, validation, supervision, writing—review and editing. All authors read and approved the manuscript.
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Communicated by Neal Stewart.
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299_2023_3050_MOESM1_ESM.rar
Supplementary file1 (RAR 207 kb) Supplementary material 1 Supplemental Materials and Methods. Supplementary material 2 sgRNA Sites and Sequences. Supplementary material 3 Statistical Data of NGS. Supplementary material 4 Results of Tissue Culture Optimization.
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Chen, Y., Xiang, H., Jia, L. et al. High-throughput creation of Nicotiana tabacum gene-targeted mutants based on CRISPR/Cas9. Plant Cell Rep 42, 2039–2042 (2023). https://doi.org/10.1007/s00299-023-03050-5
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DOI: https://doi.org/10.1007/s00299-023-03050-5