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Real-time transposable element activity in individual live cells
Journal article   Open access   Peer reviewed

Real-time transposable element activity in individual live cells

Neil H Kim, Gloria Lee, Nicholas A Sherer, K Michael Martini, Nigel Goldenfeld and Thomas E Kuhlman
Proceedings of the National Academy of Sciences - PNAS, Vol.113(26), pp.7278-7283
06/28/2016
DOI: 10.1073/pnas.1601833113
PMCID: PMC4932956
PMID: 27298350
url
https://doi.org/10.1073/pnas.1601833113View
Published (Version of record) Open Access

Abstract

The excision and reintegration of transposable elements (TEs) restructure their host genomes, generating cellular diversity involved in evolution, development, and the etiology of human diseases. Our current knowledge of TE behavior primarily results from bulk techniques that generate time and cell ensemble averages, but cannot capture cell-to-cell variation or local environmental and temporal variability. We have developed an experimental system based on the bacterial TE IS608 that uses fluorescent reporters to directly observe single TE excision events in individual cells in real time. We find that TE activity depends upon the TE's orientation in the genome and the amount of transposase protein in the cell. We also find that TE activity is highly variable throughout the lifetime of the cell. Upon entering stationary phase, TE activity increases in cells hereditarily predisposed to TE activity. These direct observations demonstrate that real-time live-cell imaging of evolution at the molecular and individual event level is a powerful tool for the exploration of genome plasticity in stressed cells.
Escherichia coli - genetics Plasmids Transposases - genetics Bacterial Proteins - genetics Fluorescence Luminescent Proteins - genetics Gene Dosage DNA Transposable Elements Genes, Reporter

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