Transcription
activation
of
genes
by
estrogens
is
driven
enhancers,
which
are
often
located
within
the
same
Topologically
Associating
Domain
(TAD)
as
non-targeted
promoters.
We
investigated
how
acute
enhancer-driven
affects
neighbouring
non-target
TAD.
Using
single-molecule
RNA
FISH
(smFISH),
we
tracked
transcription
TFF1
(enhancer-targeted)
and
TFF3
(non-targeted)
during
estrogen
stimulation.
observed
mutually
exclusive
expression
patterns:
peaked
at
1
hour,
while
reached
its
peak
3
hours,
after
’s
had
diminished.
Chromatin
looping
data
indicated
that
enhancer
loops
with
but
not
,
suggesting
upregulation
due
to
direct
enhancer-promoter
interactions.
CRISPR
deletion
1,6-hexanediol
(HD)
exposure
revealed
enhancer:promoter
undergo
Liquid-Liquid
Phase
Separation
(LLPS),
sequesters
transcriptional
machinery
inhibits
expression.
As
signalling
wanes
or
LLPS
disrupted,
declines
increases.
Our
findings
reveal
can
indirectly
influence
genes,
highlighting
a
dynamic
shift
in
gene
progresses.
FEBS Journal,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Jan. 30, 2025
Transcription,
a
crucial
step
in
the
regulation
of
gene
expression,
is
tightly
controlled
and
involves
several
essential
processes,
such
as
chromatin
organization,
recognition
specific
genomic
sequences,
DNA
binding,
ultimately
recruiting
transcriptional
machinery
to
facilitate
transcript
synthesis.
At
center
this
are
transcription
factors
(TFs),
which
comprise
at
least
one
DNA-binding
domain
(DBD)
an
effector
(ED).
Although
structure
function
DBDs
have
been
well
studied,
our
knowledge
domains
limited.
EDs
particular
importance
generating
distinct
responses
between
protein
members
same
TF
family
that
similar
specificities.
The
study
activity
conferred
by
has
traditionally
conducted
through
examining
protein-protein
interactions.
However,
recent
research
uncovered
alternative
mechanisms
regulate
formation
condensates
increase
local
concentration
factors,
cofactors,
coregulated
genes,
binding.
Here,
we
provide
comprehensive
overview
known
roles
factor
EDs,
with
focus
on
disordered
regions.
Additionally,
emphasize
significance
intrinsically
regions
(IDRs)
during
regulation.
We
examine
underlying
establishment
maintenance
specificity
structural
properties
predominantly
EDs.
then
current
understanding
these
domains,
including
their
physical
chemical
characteristics,
functional
roles.
Nature Communications,
Journal Year:
2025,
Volume and Issue:
16(1)
Published: Feb. 14, 2025
CRISPR
activation
(CRISPRa)
is
a
powerful
tool
for
endogenous
gene
activation,
yet
the
mechanisms
underlying
its
optimal
transcriptional
remain
unclear.
By
monitoring
real-time
bursts,
we
find
that
CRISPRa
modulates
both
burst
duration
and
amplitude.
Our
quantitative
imaging
reveals
CRISPR-SunTag
activators,
with
three
tandem
VP64-p65-Rta
(VPR),
form
liquid-like
condensates
exhibit
high
potency.
Although
visible
are
associated
some
RNA
overall
levels
of
phase
separation
do
not
correlate
bursting
or
strength
in
individual
cells.
When
number
SunTag
scaffolds
increased
to
10
more,
solid-like
form,
sequestering
co-activators
such
as
p300
MED1.
These
display
low
dynamicity
liquidity,
resulting
ineffective
activation.
Overall,
our
studies
characterize
various
phase-separated
systems
highlighting
foundational
principles
engineering
CRISPR-based
programmable
synthetic
appropriate
properties
effectively
modulate
expression.
The
elusive.
Here,
authors
show
activators
droplets
boost
expression,
while
creates
sequester
coactivators,
impairing
Transcription
activation
of
genes
by
estrogen
is
driven
enhancers,
which
are
often
located
within
the
same
Topologically
Associating
Domain
(TAD)
as
non-targeted
promoters.
We
investigated
how
acute
enhancer-driven
affects
neighbouring
non-target
TAD.
Using
single-molecule
RNA
FISH
(smFISH),
we
tracked
transcription
TFF1
(enhancer-targeted)
and
TFF3
(non-targeted)
during
stimulation.
observed
mutually
exclusive
expression
patterns:
peaked
at
1
hour,
while
reached
its
peak
3
hours,
after
’s
had
diminished.
Chromatin
looping
data
indicated
that
enhancer
loops
with
but
not
,
suggesting
upregulation
due
to
direct
enhancer-promoter
interactions.
CRISPR
deletion
1,6-hexanediol
(HD)
exposure
revealed
enhancer:promoter
undergo
Liquid-Liquid
Phase
Separation
(LLPS),
sequesters
transcriptional
machinery
inhibits
expression.
As
signalling
wanes
or
LLPS
disrupted,
declines
increases.
Our
findings
reveal
can
indirectly
influence
genes,
highlighting
a
dynamic
shift
in
gene
progresses.
