上海交通大学樊春海团队研制了基于DNA折纸的单分子CRISPR机器用于空间分辨搜索。相关研究成果于2022年6月23日发表在国际顶尖学术期刊《德国应用化学》。
重新利用RNA引导的核酸内切酶Cas9开发人工CRISPR分子机器代表了合成分子信息处理的一个新方向。然而，基于CRISPR-Cas9的机器的运行依赖于对自由扩散的sgRNA/Cas9的分子识别，这使得执行空间调控的局部搜索或导航具有实际挑战性。
该文中，研究人员开发了一种基于DNA折纸的单分子CRISPR机器，它可以通过自由或局部搜索模式执行空间分辨DNA切割。当以纳米级精度在DNA折纸上的特定位置触发时，自由搜索模式会导致搜索活动随距离逐渐衰减，而局部模式会产生空间受限的搜索活动。
该工作扩展了CRISPR分子机器的功能，并为开发集成分子电路和高通量核酸检测奠定了基础。
附：英文原文
Title: DNA origami-based single-molecule CRISPR machines for spatially resolved searching
Author: Yaya Hao, Mingqiang Li, Qian Zhang, Jiye Shi, Jiang Li, Qian Li, Chunhai Fan, Fei Wang
Issue&Volume: 2022-06-23
Abstract: Repurposing the RNA-guided endonuclease Cas9 to develop artificial CRISPR molecular machines represents a new direction toward synthetic molecular information processing. The operation of CRISPR-Cas9-based machines, nevertheless, relies on the molecular recognition of freely diffused sgRNA/Cas9, making it practically challenging to perform spatially regulated localized searching or navigation. Here, we develop a DNA origami-based single-molecule CRISPR machine that can perform spatially resolved DNA cleavage via either free or localized searching modes. When triggered at a specific site on the DNA origami with nanoscale accuracy, the free searching mode leads to searching activity that gradually decays with the distance, whereas the localized mode generates spatially-confined searching activity. Our work expands the function of CRISPR molecular machines and lays foundations to develop integrated molecular circuits and high-throughput nucleic acid detection.
DOI: 10.1002/anie.202205460
Source: https://onlinelibrary.wiley.com/doi/10.1002/anie.202205460
