结构可变的CRISPR/Cas12a聚 (胺) 激活剂,用于高度敏感地检测 (II) 离子
David Septian Sumanto Marpaung1, Ya-Yu Chen2, Murali Mohana Rao Singuru2
1International Ph.D. Program in Biomedical and Materials Science, Tunghai University, Taichung 407224, Taiwan; Department of Biosystems Engineering, Institut Teknologi Sumatera, Lampung Selatan 35365, Indonesia.
International journal of biological macromolecules
|July 5, 2025
概括
研究人员重新利用了CRISPR/Cas12a的目标外效应来检测. 一种新型的DNA激活剂与形成了针头,抑制了Cas12a的活性,用于敏感的环境监测.
科学领域:
- 生物技术是生物技术.
- 环境科学 环境科学
- 分子生物学分子生物学
背景情况:
- 虽然CRISPR/Cas系统很强大,但它可能会表现出非目标效应,导致意外的遗传修饰.
- 非目标活动在治疗应用中存在挑战,但为新的生物传感策略提供了潜力.
研究的目的:
- 重新利用CRISPR/Cas12a系统对离子 (Hg2+) 的敏感和选择性检测的非目标效应.
- 开发一种新的生物传感器,利用结构可变的DNA激活剂进行实时Hg2+监测.
主要方法:
- 两段聚-T DNA 激活器与 CRISPR/Cas12a 系统的集成.
- 利用提氨酸-Hg2+-提氨酸基配对诱导发针形成并调节Cas12a跨裂变活性.
- 使用循环二元化 (CD),尿素-PAGE和本地PAGE进行表征,以分析结构变化和催化效率.
主要成果:
- 增加Hg2+度促进了发针的形成,抑制了Cas12a的活动,并产生了ON-OFF光信号.
- 优化的生物传感器实现了0-30nM的线性检测范围和Hg2+的0.372nM的低检测极限.
- 对Hg2+具有很高的选择性,并在田间水样分析中成功应用.
结论:
- 该研究成功地将CRISPR/Cas12a的目标外效应用于先进的Hg2+检测,展示了环境监测中的新应用.
- 开发的生物传感器提供了一个灵敏,有选择性和强大的平台,用于检测环境样本中的重金属.
- 这项工作为利用基于CRISPR的目标外现象在开发创新的传感技术方面提供了基础.
相关概念视频
01:59CRISPR
Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced Short...
01:59CRISPR
Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced Short...
01:28CRISPR/Cas9 Genome Editing
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...