Live cell imaging of single genomic loci with quantum dot-labeled TALEs

Yingxin Ma, Mingxiu Wang, Wei Li, Zhiping Zhang, Xiaowei Zhang, Tianwei Tan, Xian-En Zhang () and Zongqiang Cui ()
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Yingxin Ma: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences
Mingxiu Wang: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences
Wei Li: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences
Zhiping Zhang: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences
Xiaowei Zhang: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences
Tianwei Tan: Beijing Key Laboratory of Bioprocess, College of Life Science and Technology, Beijing University of Chemical Technology
Xian-En Zhang: CAS Center for Biological Macromolecules, National Key Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences
Zongqiang Cui: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences

Nature Communications, 2017, vol. 8, issue 1, 1-8

Abstract: Abstract Single genomic loci are often related to specific cellular functions, genetic diseases, or pathogenic infections. Visualization of single genomic loci in live human cells is currently of great interest, yet it remains challenging. Here, we describe a strategy for live cell imaging of single genomic loci by combining transcription activator-like effectors (TALEs) with a quantum dot labelling technique. We design and select a pair of TALEs that specifically target HIV-1 proviral DNA sequences, and use bioorthogonal ligation reactions to label them with different colour quantum dots (QDs). These QD-labelled TALEs are able to enter the cell nucleus to provide fluorescent signals to identify single gene loci. Based on the co-localization of the pair of different coloured QD-labelled TALEs, we determine and map single-copy HIV-1 provirus loci in human chromosomes in live host cells.

Date: 2017
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DOI: 10.1038/ncomms15318

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