可诱导电话卡:开发用于实验控制的转子子插入的小鼠试剂in vivo
Simona Sarafinovska1,2, Arthi Venkatesan1,2, Titilope Akinwe1,2
1Department of Genetics, Washington University School of Medicine, Saint Louis, MO, USA.
bioRxiv : the preprint server for biology
|November 24, 2025
概括
我们在小鼠中开发了可诱导的呼叫卡 (iCC),以精确控制DNA记录. Jun-iCC捕获了神经活动,而Sp1-iCC显示了更高的插入,但导致了发育问题.
科学领域:
- 遗传学 遗传学 是一个
- 神经科学是一个神经科学.
- 分子生物学分子生物学
背景情况:
- PiggyBac转化酶有助于在小鼠中进行基因选.
- piggyBac与转录因子 (TF) 的融合产生呼叫卡来标记DNA结合点.
- 诱导性转体酶是必要的,以精确的时间控制的记录在体内.
研究的目的:
- 在小鼠大脑中设计和测试可诱导的呼叫卡 (iCC),用于药物诱导的DNA记录.
- 评估Jun-iCC和Sp1-iCC系统的有效性和时间控制.
- 评估可诱导的TF-转化酶融合对潜在的发育影响.
主要方法:
- 工程 knock-in 小鼠与他莫西芬诱导的TF-转化酶融合 (Jun-iCC和Sp1-iCC).
- 在体外和体内测试iCC系统,以检测插入效率和时间控制.
- 评估小鼠的发育,行为和神经活动记录.
主要成果:
- Jun-iCC小鼠在发作期间显示出塔莫西芬依赖的记录和捕获的神经活动,但插入次数较低.
- Sp1-iCC导致了更高的插入数量,但导致了发育异常,如降低活力和眼.
- ERT2域与Sp1的相互作用可能是观察到的发育缺陷的基础.
结论:
- 可诱导电话卡使药物可诱导的转子子组合 in vivo.
- Jun-iCC适用于记录短暂的神经活动,尽管插入率低.
- Sp1-iCC需要进一步优化,以减轻发育方面的副作用,同时利用其高插入能力.
相关概念视频
01:16In-vitro Mutagenesis
16.0K
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
16.0K
02:53Conservative Site-specific Recombination and Phase Variation
6.6K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
6.6K