基于胞嘧啶碱基编辑系统构建无肌间刺鱼类品系
CONSTRUCTION OF FISH WITHOUT INTERMUSCULAR BONES BASED ON CYTOSINE BASE EDITING SYSTEM
摘要
Abstract
To verify the editing effect of new cytosine base editor(CBE)in fish,this study took the runx2b that regu-lates the formation of intermuscular bones(IBs)in fish as the target gene,and explored its editing efficiency,pheno-type and related gene expression on the constructed mutation zebrafish line.The results showed that the mutation effi-ciency in the F0 generation was 60%.Skeletal staining demonstrated a 55%reduction in IBs in F0 generation mutants compared to wild-type controls.In the F1 generation produced by self-crossing of the F0 generation,only heterozygotes and homozygotes were observed,with homozygotes exhibiting completely IBs absence and stable inheritance across generations.Western Blot analysis confirmed the absence of runx2b protein in mutant,while RT-qPCR showed signifi-cantly downregulated expression of osteoblast differentiation and mineralization-related genes(entpd5a,sp7,and alpl)in mutants.The results indicate that CBE not only achieves high editing efficiency in fish,but also enables IBs free phenotypes,shortens homozygotes generation time,and expands the utility of base editing in aquatic species.This study provides a foundation for applying GE in breeding economic fish.关键词
CBE/肌间刺/runx2b/编辑效率/基因表达/斑马鱼Key words
CBE/Intermuscular bones/runx2b gene/Editing efficiency/Gene expression/Danio rerio分类
生物科学引用本文复制引用
孙秋洁,王博凯,肖正雨,徐天宏,王文秀,汪垚,高泽霞,聂春红..基于胞嘧啶碱基编辑系统构建无肌间刺鱼类品系[J].水生生物学报,2025,49(12):33-39,7.基金项目
国家自然科学基金重点项目(32330108) （32330108）
国家自然科学基金青年科学基金项目(32202917) （32202917）
国家大宗淡水鱼类产业技术体系建设项目(CARS-45-01) （CARS-45-01）
湖北省自然科学基金(2022CFB819) （2022CFB819）
湖北省技术创新计划项目(2024BBA001) （2024BBA001）
湖北农业生物基因组育种专项(2024BBB017) （2024BBB017）
中央高校基本科研业务费专项资金(2662023SCQD002)资助[Supported by the Key Program of the National Natural Science Foundation of China(32330108) （2662023SCQD002）
the Young Scientists Fund of the National Natural Science Foundation of China(32202917) （32202917）
the Earmarked Fund for Agriculture Research System of China(CARS-45-01) （CARS-45-01）
the Hubei Provincial Natural Science Foundation of China(2022CFB819) （2022CFB819）
Hubei Agricultural Gene Editing Platform Technology and Breeding R and D(2024BBA001) （2024BBA001）
the Hubei Provincial Agricultural Biology Genomic Breeding Program(2024BBB017) （2024BBB017）
the Fundamental Research Funds for the Central Universities(2662023SCQD002)] （2662023SCQD002）
