Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, 08003 Barcelona, Spain.
Universitat Pompeu Fabra (UPF), Dr Aiguader 88, 08003 Barcelona, Spain.
Nucleic Acids Res. 2018 Mar 16;46(5):e30. doi: 10.1093/nar/gkx1271.
CRISPR/dCas9-based labeling has allowed direct visualization of genomic regions in living cells. However, poor labeling efficiency and signal-to-background ratio have limited its application to visualize genome organization using super-resolution microscopy. We developed (Po)STAC (Polycistronic SunTAg modified CRISPR) by combining CRISPR/dCas9 with SunTag labeling and polycistronic vectors. (Po)STAC enhances both labeling efficiency and fluorescence signal detected from labeled loci enabling live cell imaging as well as super-resolution fixed-cell imaging of multiple genes with high spatiotemporal resolution.
基于 CRISPR/dCas9 的标记技术允许在活细胞中直接可视化基因组区域。然而，标记效率和信号背景比低限制了其应用，使其无法使用超分辨率显微镜可视化基因组结构。我们通过将 CRISPR/dCas9 与 SunTag 标记和多顺反子载体相结合，开发了 (Po)STAC（多顺反子 SunTag 修饰的 CRISPR）。(Po)STAC 提高了标记效率和从标记基因座检测到的荧光信号，使活细胞成像以及具有高时空分辨率的多个基因的超分辨率固定细胞成像成为可能。
