产生和表征一种受他莫西芬诱导的CRE驱动鼠,用于微质中的转基因表达
Elliot J Glotfelty1, Lamarque M Coke1, Evan E Hart1,2
1Intramural Research Program, National Institute on Drug Abuse, Baltimore, MD 21224.
bioRxiv : the preprint server for biology
|August 8, 2025
概括
研究人员开发了一种新的转基因大鼠模型,用于研究中枢神经系统的免疫细胞 - - 微质细胞. 这种工具可以精确地对大鼠的微质进行基因操纵,从而促进对大脑功能和疾病的研究.
科学领域:
- 神经科学是一个神经科学.
- 免疫学 免疫学 免疫学
- 遗传学 是一个遗传学.
背景情况:
- 微质细胞是中枢神经系统 (CNS) 中的关键免疫细胞,具有多样化的作用.
- 现有的微质细胞研究的转基因工具主要在小鼠身上.
- 鼠对研究微质功能,特别是复杂行为的研究具有优势.
研究的目的:
- 创建和验证一种新的转基因大鼠模型,用于诱导和特定的微质基因操纵.
- 为了实现对小鼠微质和外周免疫细胞功能的先进研究.
主要方法:
- 在微质分类素受体 (Cx3cr1) 促进剂下开发了一种表达他莫西芬诱导的Cre复合酶 (CreERT2) 的BAC转基因大鼠.
- 通过与记者大鼠交叉并评估Iba1局部化,确认了微质特异性.
- 利用流细胞计分析Cx3cr1+细胞在tamoxifen治疗后的组织中的重组.
主要成果:
- 成功生成了Cx3cr1-CreERT2转基因大鼠.
- 已证明可以诱导他莫西芬和微质细胞的Cre表达.
- 证实这些老鼠与野生类型对照相比表现出正常的认知行为.
- 在微质细胞和外围免疫细胞中显示了时间和Cre依赖的重组.
结论:
- 已经建立了一个新的转基因大鼠模型用于微质细胞研究.
- 这种模型允许精确的,可诱导的基因向小质在老鼠.
- 该模型适用于研究中枢神经系统和外周系统中微质的作用.
相关概念视频
00:53Transgenic Organisms
Overview
01:16In-vitro Mutagenesis
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.
02:53Conservative Site-specific Recombination and Phase Variation
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...
02:11Reporter Genes
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Commonly used reporter...