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使用基于CRISPR/CAS9的基因向方式,生成WT1-td番茄敲进的Cynomolgus子胚胎干细胞系,WT1-205的基因向

Ahjol Hyraht¹, Fenglin Zhan¹, Rongrong Guo¹

¹State Key Laboratory of Primate Biomedical Research, Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China.

Stem cell research

|June 4, 2025

概括

研究人员在Cynomolgus子中创建了一个WT1 (威尔姆斯瘤1) 记者干细胞系. 这个工具跟踪细胞发展成脏和生殖腺,推进发育生物学研究.

科学领域:

发育生物学发展生物学
干细胞生物学干细胞生物学
遗传学是一个遗传学.

背景情况:

威尔姆斯瘤1 (WT1) 基因对于脏,生殖腺和上腺皮层等中间结构的发展至关重要.
了解WT1基因功能对于研究器官生成和发育异常至关重要.

研究的目的:

为了生成一个WT1记者 cynomolgus胚胎干细胞 (cyESC) 线来跟踪WT1表达细胞.
为了验证本报告员线在研究中介介皮层发育中的实用性.

主要方法:

使用CRISPR/Cas9技术,在cyESCs中的WT1停止编码子中敲入tdTomato记者基因.
一个PGK-Neo选择磁带 (LSL) 被包括在内,随后被Cre recombinase切除.
实验室差异化协议被用于产生和淋巴状细胞.

主要成果:

在分化细胞中观察到番茄光,表明成功向WT1基因.
与已知的标记物 (淋巴腺GATA4,脏PAX2) 同定位tdTomato证实了报告者的特异性.
阳性WT1细胞表达了其他相关的中间介质皮质基因.

结论:

生成的WT1-tdTomato cyESC线是研究非人类灵长类动物中WT1-驱动的发育过程的宝贵工具.
这种记者线有助于研究介质皮层发育和细胞命运的确定.
这些发现有助于更好地了解灵长类动物胚胎发生和再生医学中的潜在应用.

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