美国斯坦福大学亓磊课题组开发出CRISPR介导的基因组编辑和转录的实时成像工具。该项研究成果2019年9月5日在线发表在《科学》上。

他们报告了一种强大的通用工具，名为CRISPR活细胞荧光原位杂交（LiveFISH）技术，可使用荧光寡核苷酸在包括原代细胞在内的广泛细胞类型中进行基因组追踪。

CRISPR指导RNA的内在稳定性转换使得LiveFISH能够准确地检测染色体疾病，例如产前羊水细胞中的Patau综合征，并示踪人T淋巴细胞中的多个基因座。此外，LiveFISH示踪由CRISPR-Cas9介导的编辑和随后的染色体易位导致的DNA双链断裂的实时动态。 最后，结合Cas9和Cas13系统，LiveFISH可以同时显示活细胞中的基因组DNA和RNA转录本。LiveFISH方法可在单细胞的基因组编辑，转录和重排过程中实现DNA和RNA的实时成像。

附：英文原文

Title: CRISPR-mediated live imaging of genome editing and transcription

Author: Haifeng Wang, Muneaki Nakamura, Timothy R. Abbott, Dehua Zhao, Kaiwen Luo, Cordelia Yu, Cindy M. Nguyen, Albert Lo, Timothy P. Daley, Marie La Russa, Yanxia Liu, Lei S. Qi

Issue&Volume: 2019/09/05

Abstract: We report a robust, versatile approach named CRISPR Live-cell fluorescent in situ hybridization (LiveFISH) using fluorescent oligos for genome tracking in broad cell types including primary cells. An intrinsic stability switch of CRISPR guide RNAs enables LiveFISH to accurately detect chromosomal disorders such as Patau Syndrome in prenatal amniotic fluid cells and track multiple loci in human T lymphocytes. In addition, LiveFISH tracks the real-time movement of DNA double-strand breaks induced by CRISPR-Cas9-mediated editing and consequent chromosome translocations. Finally, combining Cas9 and Cas13 systems, LiveFISH allows for simultaneous visualization of genomic DNA and RNA transcripts in living cells. The LiveFISH approach enables real-time live imaging of DNA and RNA during genome editing, transcription, and rearrangements in single cells.

DOI: 10.1126/science.aax7852

Source:https://science.sciencemag.org/content/early/2019/09/04/science.aax7852