Multiplexed Dark FRET Biosensors: An accessible live-cell platform for target- and cell-specific monitoring of protein-protein interactions in 2D and 3D model systems
Published: May 12, 2025
Abstract
Simultaneously
monitoring
multiple
protein-protein
interactions
in
live
cells
remains
a
key
challenge
biology
and
drug
discovery.
While
multiplexed
FRET
enables
parallel
molecular
readouts,
existing
approaches
are
often
constrained
by
spectral
overlap,
complex
instrumentation,
or
incompatibility
with
live-cell
models.
To
overcome
these
limitations
increase
accessibility
to
the
broader
biological
community,
we
present
Multiplexed
Dark
(MDF),
genetically
encoded
platform
that
uses
spectrally
distinct
donors
(mNeonGreen,
mScarlet-I3)
paired
non-emissive
acceptors
(ShadowY,
ShadowR).
Using
fluorescence
lifetime
detection,
demonstrate
MDF’s
versatility
through
three
biologically
translationally
relevant
examples:
(1)
cell-type
specific
biosensing
organoids,
as
exemplified
3D
neuro–glial
spheroids;
(2)
target
specificity
for
discovery,
discrimination
of
TNFR1
versus
TNFR2
receptor
conformations;
(3)
protein
misfolding,
simultaneous
alpha-synuclein
oligomerization
misfolding.
MDF
provides
scalable
framework
real-time,
across
high-throughput,
target-specific,
tissue-level
applications
systems.
Language: Английский
Association Between Serum α-Synuclein Levels and Neurocognitive Deficits in Patients with Obstructive Sleep Apnea: A Case-Control Study
Sleep Medicine,
Journal Year:
2025,
Volume and Issue:
unknown, P. 106583 - 106583
Published: May 1, 2025
Language: Английский