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
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.