Live visualization of genomic loci with BiFC-TALE

Sci Rep. 2017 Jan 11:7:40192. doi: 10.1038/srep40192.

Abstract

Tracking the dynamics of genomic loci is important for understanding the mechanisms of fundamental intracellular processes. However, fluorescent labeling and imaging of such loci in live cells have been challenging. One of the major reasons is the low signal-to-background ratio (SBR) of images mainly caused by the background fluorescence from diffuse full-length fluorescent proteins (FPs) in the living nucleus, hampering the application of live cell genomic labeling methods. Here, combining bimolecular fluorescence complementation (BiFC) and transcription activator-like effector (TALE) technologies, we developed a novel method for labeling genomic loci (BiFC-TALE), which largely reduces the background fluorescence level. Using BiFC-TALE, we demonstrated a significantly improved SBR by imaging telomeres and centromeres in living cells in comparison with the methods using full-length FP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Genetic Loci*
  • Humans
  • Intravital Microscopy / methods*
  • Microscopy, Fluorescence / methods*
  • Staining and Labeling / methods*