Department of Biomedical Engineering, College of Engineering, Peking University, Beijing 100871, China.
School of Life Sciences, Peking University, Beijing 100871, China.
Nucleic Acids Res. 2019 Nov 18;47(20):e131. doi: 10.1093/nar/gkz752.
Clustered regularly interspaced short palindromic repeats (CRISPR)-based genomic imaging systems predominantly rely on fluorescent protein reporters, which lack the optical properties essential for sensitive dynamic imaging. Here, we modified the CRISPR single-guide RNA (sgRNA) to carry two distinct molecular beacons (MBs) that can undergo fluorescence resonance energy transfer (FRET) and demonstrated that the resulting system, CRISPR/dual-FRET MB, enables dynamic imaging of non-repetitive genomic loci with only three unique sgRNAs.
基于成簇规律间隔短回文重复序列（CRISPR）的基因组成像系统主要依赖于荧光蛋白报告基因，而这些报告基因缺乏进行灵敏动态成像所需的光学特性。在这里，我们对 CRISPR 单指导 RNA（sgRNA）进行了修饰，使其携带两个不同的分子信标（MB），这两个分子信标可以进行荧光共振能量转移（FRET），并证明了所得到的系统，即 CRISPR/双重-FRET MB，可以仅使用三个独特的 sgRNA 对非重复基因组基因座进行动态成像。
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