Nanog Fluctuations in Embryonic Stem Cells Highlight the Problem of Measurement in Cell Biology.

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.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Smith RCG,Stumpf PS,Ridden SJ,Sim A,Filippi S,Harrington HA,MacArthur BD

doi

10.1016/j.bpj.2017.05.005

subject

Has Abstract

pub_date

2017-06-20 00:00:00

pages

2641-2652

issue

12

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(17)30509-X

journal_volume

112

pub_type

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