Open AccessLive cell imaging of single genomic loci with quantum dot-labeled TALEs
Author(s) -
Yingxin Ma,
Mingxiu Wang,
Wei Li,
Zhiping Zhang,
Xiaowei Zhang,
Tianwei Tan,
Xian-En Zhang,
Zongqiang Cui
Publication year - 2017
Publication title -
nature communications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.559
H-Index - 365
ISSN - 2041-1723
DOI - 10.1038/ncomms15318
Subject(s) - provirus , biology , genetics , live cell imaging , single cell analysis , quantum dot , dna , human genome , gene , genomic dna , computational biology , genome , cell , physics , quantum mechanics
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.