26
IRUS Total
Downloads
  Altmetric

Nanog fluctuations in embryonic stem cells highlight the problem of Measurement in cell biology

File Description SizeFormat 
mmc2.pdfPublished version9.91 MBAdobe PDFView/Open
Title: Nanog fluctuations in embryonic stem cells highlight the problem of Measurement in cell biology
Authors: Smith, RCG
Stumpf, PS
Ridden, SJ
Sim, A
Filippi, SL
Harrington, HA
MacArthur, BD
Item Type: Journal Article
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.
Issue Date: 20-Jun-2017
Date of Acceptance: 5-May-2017
URI: http://hdl.handle.net/10044/1/48537
DOI: https://dx.doi.org/10.1016/j.bpj.2017.05.005
ISSN: 1542-0086
Publisher: Biophysical Society
Start Page: 2641
End Page: 2652
Journal / Book Title: Biophysical Journal
Volume: 112
Issue: 12
Copyright Statement: © 2017 Biophysical Society. This is an open access article under the CC BY license (http:// creativecommons.org/licenses/by/4.0/).
Keywords: Science & Technology
Life Sciences & Biomedicine
Biophysics
STOCHASTIC GENE-EXPRESSION
GREEN-FLUORESCENT PROTEIN
GROUND-STATE
SELF-RENEWAL
POSITIVE-FEEDBACK
LIVING CELLS
SINGLE-CELL
PLURIPOTENCY
HETEROGENEITY
NETWORKS
02 Physical Sciences
03 Chemical Sciences
06 Biological Sciences
Appears in Collections:School of Public Health