Xue Chenxiao, Qiu Fengti, Wang Yuxiang, Li Boshu, Zhao Kevin Tianmeng, Chen Kunling, Gao Caixia
State Key Laboratory of Plant Cell and Chromosome Engineering, Center for Genome Editing, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
College of Advanced Agricultural Sciences, University of Chinese Academy of Sciences, Beijing, China.
Nat Biotechnol. 2023 Dec;41(12):1758-1764. doi: 10.1038/s41587-023-01707-w. Epub 2023 Mar 9.
The ability to control gene expression and generate quantitative phenotypic changes is essential for breeding new and desired traits into crops. Here we report an efficient, facile method for downregulating gene expression to predictable, desired levels by engineering upstream open reading frames (uORFs). We used base editing or prime editing to generate de novo uORFs or to extend existing uORFs by mutating their stop codons. By combining these approaches, we generated a suite of uORFs that incrementally downregulate the translation of primary open reading frames (pORFs) to 2.5-84.9% of the wild-type level. By editing the 5' untranslated region of OsDLT, which encodes a member of the GRAS family and is involved in the brassinosteroid transduction pathway, we obtained, as predicted, a series of rice plants with varied plant heights and tiller numbers. These methods offer an efficient way to obtain genome-edited plants with graded expression of traits.
控制基因表达并产生定量表型变化的能力对于将新的所需性状培育到作物中至关重要。在此，我们报告了一种高效、简便的方法，通过改造上游开放阅读框（uORF）将基因表达下调至可预测的所需水平。我们使用碱基编辑或引导编辑来从头生成uORF，或通过突变其终止密码子来扩展现有uORF。通过结合这些方法，我们生成了一套uORF，它们将主要开放阅读框（pORF）的翻译逐步下调至野生型水平的2.5% - 84.9%。通过编辑编码GRAS家族成员且参与油菜素内酯信号转导途径的OsDLT的5'非翻译区，我们如预期获得了一系列具有不同株高和分蘖数的水稻植株。这些方法为获得具有性状分级表达的基因组编辑植物提供了一种有效途径。
