Author
Smith, R
Stumpf, P
Ridden, S
Sim, A
Filippi, S
Harrington, H
MacArthur, B
Journal title
Biophysical Journal
DOI
10.1016/j.bpj.2017.05.005
Issue
12
Volume
112
Last updated
2024-03-21T23:13:59.073+00:00
Page
2641-2652
Abstract
A number of important pluripotency regulators, including the transcription factor Nanog, are observed to fluctuate stochastically in individual embryonic stem cells. By transiently priming cells for commitment to different lineages, these fluctuations are thought to be important to the maintenance of, and exit from, pluripotency. However, because temporal changes in intracellular protein abundances cannot be measured directly in live cells, fluctuations are typically assessed using genetically engineered reporter cell lines that produce a fluorescent signal as a proxy for protein expression. Here, using a combination of mathematical modeling and experiment, we show that there are unforeseen ways in which widely used reporter strategies can systematically disturb the dynamics they are intended to monitor, sometimes giving profoundly misleading results. In the case of Nanog, we show how genetic reporters can compromise the behavior of important pluripotency-sustaining positive feedback loops, and induce a bifurcation in the underlying dynamics that gives rise to heterogeneous Nanog expression patterns in reporter cell lines that are not representative of the wild-type. These findings help explain the range of published observations of Nanog variability and highlight the problem of measurement in live cells.
Symplectic ID
694272
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Publication type
Journal Article
Publication date
20 Jun 2017
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