利用CRISPR/Cas9基因编辑技术构建斑马鱼map3k15敲除品系

doi:10.16352/j.issn.1001-6325.2025.03.0290

基础医学与临床 ›› 2025, Vol. 45 ›› Issue (3): 290-297.doi: 10.16352/j.issn.1001-6325.2025.03.0290

利用CRISPR/Cas9基因编辑技术构建斑马鱼map3k15敲除品系

贾淳钰¹, 沈延², 陈罡¹, 郑可¹, 秦岩^1*, 李雪梅¹

1.中国医学科学院北京协和医学院北京协和医院肾内科,北京 100730;
2.北京大学生命科学学院细胞增殖与分化教育部重点实验室,北京 100871

收稿日期:2024-11-07 修回日期:2024-12-25 发布日期:2025-02-25
通讯作者: ^*qinyanbeijing@126.com
基金资助:
国家自然科学基金(81970621)

Establishment of a zebrafish map3k15knockout line using CRISPR/Cas9 gene editing technology

JIA Chunyu¹, SHEN Yan², CHEN Gang¹, ZHENG Ke¹, QIN Yan^1*, LI Xuemei¹

1. Department of Nephrology, Peking Union Medical College Hospital, CAMS & PUMC, Beijing 100730;
2. Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing 100871, China

Received:2024-11-07 Revised:2024-12-25 Published:2025-02-25

摘要/Abstract

摘要： 目的利用CRISPR/Cas9基因编辑技术构建斑马鱼map3k15敲除纯合品系,为进一步研究map3k15在肾脏疾病方面的作用提供动物模型。方法 1)分析map3k15在斑马鱼中的表达模式;2)利用CRISPR/Cas9基因编辑技术构建斑马鱼map3k15敲除纯合品系;3)观察map3k15缺失对斑马鱼表型的影响。结果 1)map3k15在斑马鱼前肾中表达,且MAP3K15/Map3k15蛋白在多物种间具有高度保守性;2)成功构建了斑马鱼map3k15^-/-突变体,并保留+2 bp及+1 bp 2种突变品系;3)map3k15突变斑马鱼在胚胎发育过程中出现卵黄囊、心包及头部的水肿,且随发育时间延长,症状逐渐加重。结论成功构建了map3k15敲除的纯合斑马鱼品系,为未来研究map3k15在肾脏发育及疾病中的作用提供了重要的模型。

关键词: map3k15, CRISPR/Cas9基因编辑技术, 斑马鱼, 肾脏发育

Abstract: Objective To establish a homozygous zebrafish map3k15 knockout line using CRISPR/Cas9 gene editing technology so to provide an animal model for investigation of potential role of map3k15 in renal diseases. Methods 1)Analyzing the expression pattern of map3k15 in zebrafish; 2)Establishing a homozygous map3k15 knockout zebrafish line using CRISPR/Cas9; 3)Observing the phenotypic effects of map3k15 deficiency in zebrafish. Results 1)map3k15 was expressed in the zebrafish pronephros, and map3k15/map3k15 protein was found highly conserved across species; 2)map3k15^-/- mutant zebrafish model was constructed with +2 bp mutation and +1 bp mutation; 3)map3k15 mutant zebrafish exhibited edema in yolk sac, pericardium and head during embryonic development. Conclusions A homozygous zebrafish map3k15 knockout line is successfully developed which may provide an important model for future research on the role of map3k15 in renal development and disease.

Key words: map3k15, CRISPR/Cas9 gene editing technology, zebrafish, kidney development

中图分类号:

S941.42+7

贾淳钰, 沈延, 陈罡, 郑可, 秦岩, 李雪梅. 利用CRISPR/Cas9基因编辑技术构建斑马鱼map3k15敲除品系[J]. 基础医学与临床, 2025, 45(3): 290-297.

JIA Chunyu, SHEN Yan, CHEN Gang, ZHENG Ke, QIN Yan, LI Xuemei. Establishment of a zebrafish map3k15knockout line using CRISPR/Cas9 gene editing technology[J]. Basic & Clinical Medicine, 2025, 45(3): 290-297.

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利用CRISPR/Cas9基因编辑技术构建斑马鱼map3k15敲除品系

Establishment of a zebrafish map3k15knockout line using CRISPR/Cas9 gene editing technology

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