Abstract
Key message
A high-efficiency protoplast transient system was devised to screen genome editing elements in Salvia miltiorrhiza.
Abstract
Medicinal plants with high-value pharmaceutical ingredients have attracted research attention due to their beneficial effects on human health. Cell wall-free protoplasts of plants can be used to evaluate the efficiency of genome editing mutagenesis. The capabilities of gene editing in medicinal plants remain to be fully explored owing to their complex genetic background and shortfall of suitable transformation. Here, we took the Salvia miltiorrhiza as a representative example for develo** a method to screen favorable gene editing elements with high editing efficiency in medical plants by a PEG-mediated protoplast transformation. Results indicated that using the endogenous SmU6.1 of S. miltiorrhiza to drive sgRNA and the plant codon-optimized Cas9 driven by the promoter SlEF1α can enhance the efficiency of editing. In summary, we uncover an efficacious transient method for screening editing elements and shed new light on increasing gene editing efficiency in medicinal plants.
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This work was funded by China Academy of Chinese Medical Sciences (CACMS) Innovation Fund (CI2021A041003), The Ability Establishment of Sustainable Use for Valuable Chinese Medicine Resources (2060302), National Key R&D Program of China (2018YFA0900600), Shanghai Jiao Tong University (SJTU) Transmed Awards Research (20190104) and the Bill & Melinda Gates Foundation (grant numbers: OPP1199872 and INV-027291). Under the grant conditions of the Foundation, a Creative Commons Attribution 4.0 Generic License has already been assigned to the Author Accepted Manuscript version that might arise from this submission.
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JS, HZ, and KT designed the project; JS, BP, YZ, and XY performed most of the experiments; XY, XH, XF, and LL analyzed the data; JS and BP wrote this research manuscript. All the authors have read and approved the manuscript.
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Shao, J., Peng, B., Zhang, Y. et al. A high-efficient protoplast transient system for screening gene editing elements in Salvia miltiorrhiza. Plant Cell Rep 43, 45 (2024). https://doi.org/10.1007/s00299-023-03134-2
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DOI: https://doi.org/10.1007/s00299-023-03134-2