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用斑马鱼 hand2 来标记血统的Cre和CreERT2重组酶是CRISPR knock-ins

Zhitao Ming¹, Fang Liu¹, Hannah R Moran²

¹Department of Genetics, Development and Cell Biology, Iowa State University, Ames, Iowa 50011-1101 USA.

bioRxiv : the preprint server for biology

|December 16, 2024

概括

使用CRISPR-Cas9的新斑马鱼工具可以在发育过程中精确追踪hand2基因表达的血统. 这些驱动因素揭示了关于中皮和神经细胞对各种组织和血管系统的贡献的新见解.

科学领域:

发育生物学是发展生物学.
遗传学遗传学是一个
斑马鱼模型的模型

背景情况:

克里斯普尔-Cas9技术可以生成内源的克里重组酶驱动器,用于谱系追踪.
内生斑马鱼手2 Cre和CreERT2驱动程序是使用GeneWeld CRISPR-Cas9精确目标集成创建的.

研究的目的:

为了生成和表征新的内源斑马鱼手2 Cre和CreERT2驱动器.
在斑马鱼发育过程中利用这些驱动因素来追踪血统和理解手2祖先的贡献.

主要方法:

克里斯普尔-Cas9敲门技术用于产生内源的CRE驱动器.
基因精确目标集成.
交叉手2-Cre/CreERT2敲门机与基于loxP的交换机记者.
时间诱导CreERT2活动.

主要成果:

在心脏,胸,喉弧,肝脏,肠道和间皮组织,以及肠道神经元中介皮和神经谱系的广泛标记.
在静脉血管中观察到的新手2血统追踪模式.
时间分析显示,晚期出现的hand2祖先有助于静脉和肠道血管结构,幼虫诱导限制了标记特定的间皮系.

结论:

从侧板介质皮和外皮的 hand2 的祖先有助于不同的胚胎血统.
后来的手2祖先在幼虫中显示出有限的血统贡献.
开发的hand2Cre和CreERT2驱动器是研究斑马鱼胚胎和幼虫器官生成中的hand2血统的宝贵工具.

关键词:

克里斯普尔-Cas9是什么意思基因接基因接基因接在Hand2的血统中.早期和晚期出现的静脉祖先.细菌层细菌层细菌层敲进来 - 敲进来神经峰的神经峰有机体的产生有机体的产生斑马鱼是一种斑马鱼.

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