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Visualization of repetitive DNA sequences in human chromosomes with transcription activator-like effectors

Author:
Ma, Hanhui, Reyes-Gutierrez, Pablo, Pederson, Thoru
Source:
Proceedings of the National Academy of Sciences of the United States of America 2013 v.110 no.52 pp. 21048-21053
ISSN:
0027-8424
Subject:
DNA, correlation, flow cytometry, fluorescent proteins, genome, humans, image analysis, nucleotide sequences, osteosarcoma, telomeres
Abstract:
We describe a transcription activator-like effector (TALE)-based strategy, termed “TALEColor,” for labeling specific repetitive DNA sequences in human chromosomes. We designed TALEs for the human telomeric repeat and fused them with any of numerous fluorescent proteins (FPs). Expression of these TALE–telomere–FP fusion proteins in human osteosarcoma's (U2OS) cells resulted in bright signals coincident with telomeres. We also designed TALEs for centromeric sequences unique to certain chromosomes, enabling us to localize specific human chromosomes in live cells. Meanwhile we generated TALE–FPs in vitro and used them as probes to detect telomeres in fixed cells. Using human cells with different average telomere lengths, we found that the TALEColor signals correlated positively with telomere length. In addition, suspension cells were followed by imaging flow cytometry to resolve cell populations with differing telomere lengths. These methods may have significant potential both for basic chromosome and genome research as well as in clinical applications.
Agid:
1784942