相关实验视频
Updated: Jun 14, 2025
08:25
Identification of Circular RNAs using RNA Sequencing
Published on: November 14, 2019
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使用低输入RNA的全长circRNA测序方法,并在纳米孔平台上的MPTP-PD小鼠中对circRNA进行分析
Ying Wang1, Xiaohan Li1,2, Wenxiang Lu1
1State Key Lab of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, Jiangsu, 210096, China. whitecf@seu.edu.cn.
The Analyst
|September 6, 2024
概括
研究人员开发了一种新的纳米孔测序方法,从低输入样本中检测全长圆形RNA (circRNAs). 这一突破使得帕金森病模型中的全面circRNA分析成为可能,揭示出异形水平表达变化.
科学领域:
- 基因组学就是基因组学.
- 分子生物学分子生物学
- 生物信息学是一种生物信息学.
背景情况:
- 准确的全长 (FL) 转录信息对于功能分析至关重要.
- 在FL转录研究中,第三代测序 (TGS) 优先于短读测序.
- 现有的FL circRNA测序方法需要大量的RNA输入,限制它们在低生物质样本中的使用.
研究的目的:
- 利用纳米孔平台开发一种有效的方法,从低输入总RNA样本 (1-100 ng) 中检测FL circRNAs.
- 通过结合样本条形码来减少成本和运行时间.
- 在帕金森病 (PD) 鼠标模型中描述circRNA表达.
主要方法:
- 开发了一种基于纳米孔的新型测序策略,用于从总RNA的纳米克级检测FL circRNA.
- 嵌入 24 nt 条形码,用于多重复和降低成本.
- 应用该方法来对PD小鼠模型的条纹体,海马体和大脑皮层中的circRNA表达进行分析.
主要成果:
- 该方法成功地从总RNA中只需1 ng的FL circRNAs中识别出.
- 在PD和健康小鼠中确定了超过263,934个circRNA,其中很大一部分与PD相关的基因有关.
- 在异形水平上观察到circRNA表达模式的显著变化,并预测了蛋白质翻译证据.
结论:
- 开发了一种有效且具有成本效益的方法,用于从低输入样本中表征FL circRNA.
- 在异形水平上提供了对PD中circRNAs生物功能的全面见解.
- 该方法有助于更深入地了解circRNA在复杂疾病中的作用.
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