华北农学报2026，Vol.41Issue(1)：39-48,10.DOI:10.7668/hbnxb.20195725
茄科作物高效双碱基编辑新工具的开发与应用
Development and Application of a High-Efficiency Dual-Base Editing Tool for Solanaceous Crops
摘要
Abstract
This study seeks to develop a highly efficient dual-base editing tool for solanaceous crops to address the challenges of poor compatibility and low editing efficiency associated with existing base editing systems in these plants.The research involved fusing the nickase Cas9 enzyme(nCas9),which has partially lost its nuclease function,with components such as the highly efficient adenine deaminase ABE8e,the optimized cytosine deaminase sdd7-mini,and the uracil glycosylation inhibitor UGI.Systematic optimization was conducted based on the promoter preferences and co-don usage frequencies specific to solanaceous crops.Ultimately,the dual-base editor TPDBE-S was constructed.In tran-sient transformation experiments using tomato and eggplant protoplasts,this editor exhibited exceptionally high editing ef-ficiency.In the stable transformation system of tobacco,targeting the PDS gene verification revealed that four out of five positive transgenic families exhibited pronounced albinism phenotypes,thereby confirming the stable editing capability.The editing window of this editor aligns closely with the characteristics of ABE8e and sdd7-mini.Furthermore,optimizing codon preference and promoter activity in Solanaceae crops significantly enhances editing efficiency and adaptability.The construction of TPDBE-S employs a modular design.The multi-target expression cassette,based on the tRNA self-cleaving mechanism,can be rapidly assembled through one-step gene synthesis,which is both cost-effective and straight-forward.This tool addresses the gap in double-base editing specifically for Solanaceous crops.关键词
茄子/基因编辑/双碱基编辑/番茄/编辑效率Key words
Eggplant/Gene editing/Dual-base editing/Tomato/Editing efficiency分类
农业科技引用本文复制引用
田鹏,吴志明,李亚栋,张敬敬,田哲娟,康忱,高秀瑞,李冰,刘伟,武彦荣..茄科作物高效双碱基编辑新工具的开发与应用[J].华北农学报,2026,41(1):39-48,10.基金项目
河北省农林科学院博士基金项目(C23R0803) （C23R0803）
河北省自然科学基金项目(C2023301109) （C2023301109）
河北省农林科学院现代农业科技创新专项课题项目(2022KJCXZX-JZS-2) （2022KJCXZX-JZS-2）
河北省省级科技计划资助项目(21326309D) （21326309D）
