Frontiers Science Center for Transformative Molecules, Joint International Research Laboratory of Metabolic and Developmental Sciences, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China.
State Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
Plant Cell Rep. 2024 Jan 23;43(2):45. doi: 10.1007/s00299-023-03134-2.
A high-efficiency protoplast transient system was devised to screen genome editing elements in Salvia miltiorrhiza. 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 developing 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.
我们设计了一种高效的原生质体瞬时表达系统，用于筛选丹参中的基因组编辑元件。具有高价值药用成分的药用植物因其对人类健康的有益作用而引起了研究关注。无细胞壁的植物原生质体可用于评估基因组编辑诱变的效率。由于药用植物遗传背景复杂，转化方法不合适，其基因编辑的能力仍有待充分探索。在这里，我们以丹参为例，开发了一种通过 PEG 介导的原生质体转化筛选具有高效编辑能力的有利基因编辑元件的方法。结果表明，利用丹参内源性 SmU6.1 驱动 sgRNA 以及由 SlEF1α 启动子驱动的植物密码子优化的 Cas9，可以提高编辑效率。总之，我们揭示了一种有效的瞬时筛选编辑元件的方法，为提高药用植物的基因编辑效率提供了新的思路。
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