Synthetic
and
chimeric
receptors
capable
of
recognizing
responding
to
user-defined
antigens
have
enabled
“smart”
therapeutics
based
on
engineered
cells.
These
cell
engineering
tools
depend
antigen
sensors
which
are
most
often
derived
from
antibodies.
Advances
in
the
de
novo
design
proteins
protein
binders
with
potential
target
epitopes
unique
properties
faster
production
timelines
compared
Building
upon
our
previous
work
combining
a
-designed
minibinder
Spike
SARS-CoV-2
synthetic
receptor
synNotch
(SARSNotch),
we
investigated
whether
minibinders
can
be
readily
adapted
diversity
tools.
We
show
that
LCB1
easily
generalizes
next-generation
proteolytic
SNIPR
performs
similarly
previously
reported
SARSNotch.
LCB1-SNIPR
successfully
enables
detection
live
SARS-CoV-2,
an
improvement
over
SARSNotch
only
detect
cell-expressed
Spike.
To
test
generalizability
diverse
applications,
tested
as
sensor
for
(CAR).
LCB1-CAR
CD8+
T
cells
cytotoxically
Spike-expressing
further
demonstrate
two
other
directed
against
clinically
relevant
epidermal
growth
factor
able
drive
CAR-dependent
cytotoxicity
efficacy
similar
or
better
than
existing
antibody-based
CAR.
Our
findings
suggest
represent
novel
class
dramatically
expand
sensing
repertoire
Cell Systems,
Год журнала:
2022,
Номер
13(12), С. 950 - 973
Опубликована: Дек. 1, 2022
To
elucidate
principles
operating
in
native
biological
systems
and
to
develop
novel
biotechnologies,
synthetic
biology
aims
build
integrate
gene
circuits
within
transcriptional
networks.
The
utility
of
for
cell
engineering
relies
on
the
ability
control
expression
all
constituent
transgene
components.
Transgene
silencing,
defined
as
loss
over
time,
persists
an
obstacle
primary
cells
stem
with
transgenic
cargos.
In
this
review,
we
highlight
challenge
that
silencing
poses
robust
mammalian
cells,
outline
potential
molecular
mechanisms
present
approaches
preventing
silencing.
We
conclude
a
perspective
identifying
future
research
directions
improving
performance
circuits.
ABSTRACT
Cell-cell
interactions
are
central
to
development,
but
exploring
how
a
change
in
any
given
cell
relates
changes
the
neighbour
of
that
can
be
technically
challenging.
Here,
we
review
recent
developments
synthetic
biology
and
image
analysis
helping
overcome
this
problem.
We
highlight
opportunities
presented
by
these
advances
discuss
limitations
applying
them
developmental
model
systems.
The EMBO Journal,
Год журнала:
2024,
Номер
43(18), С. 4110 - 4135
Опубликована: Июль 12, 2024
Cell
communication
coordinates
developmental
processes,
maintains
homeostasis,
and
contributes
to
disease.
Therefore,
understanding
the
relationship
between
cells
in
a
shared
environment
is
crucial.
Here
we
introduce
Positive
Ultra-bright
Fluorescent
Fusion
For
Identifying
Neighbours
(PUFFFIN),
cell
neighbour-labelling
system
based
upon
secretion
uptake
of
positively
supercharged
fluorescent
protein
s36GFP.
We
fused
s36GFP
mNeonGreen
or
HaloTag,
facilitating
ultra-bright,
sensitive,
colour-of-choice
labelling.
Secretor
transfer
PUFFFIN
neighbours
while
retaining
nuclear
mCherry,
making
identification,
isolation,
investigation
live
straightforward.
can
be
delivered
cells,
tissues,
embryos
on
customisable
single-plasmid
construct
composed
interchangeable
components
with
option
incorporate
any
transgene.
This
versatility
enables
manipulation
properties,
simultaneously
labelling
surrounding
culture
vivo.
use
ask
whether
pluripotent
adjust
pace
differentiation
synchronise
their
during
exit
from
naïve
pluripotency.
offers
simple,
approach
profile
non-cell-autonomous
responses
natural
induced
changes
identity
behaviour.
ABSTRACT
In
the
developing
mouse
ventral
spinal
cord,
HES5,
a
transcription
factor
downstream
of
Notch
signalling,
is
expressed
as
evenly
spaced
clusters
high
HES5-expressing
neural
progenitor
cells
along
dorsoventral
axis.
While
signalling
requires
direct
membrane
contact
for
its
activation,
we
have
previously
shown
mathematically
that
needs
to
extend
beyond
neighbouring
HES5
pattern
emerge.
However,
presence
cellular
structures
could
enable
such
long-distance
was
unclear.
Here,
report
protrusions
are
present
all
apicobasal
axis
individual
cells.
Through
live
imaging,
show
these
dynamically
and
retract
reaching
lengths
up
∼20
µm,
enough
adjacent
The
ligand
DLL1
found
colocalise
with
protrusions,
further
supporting
idea
can
be
transduced
at
distance.
effect
on
tested
by
reducing
density
using
CDC42
inhibitor
ML141,
leading
tendency
decrease
distance
between
cell
clusters.
this
not
significant
leaves
an
open
question
about
their
role
in
fine-grained
organisation
neurogenesis.
ACS Synthetic Biology,
Год журнала:
2025,
Номер
unknown
Опубликована: Май 6, 2025
Historically,
studying
the
development
of
brain
and
central
nervous
system
(CNS)
tissues
has
been
challenging.
Human
pluripotent
stem
cell
(hPSC)
technology
allowed
for
in
vitro
reconstitution
relevant,
early
trajectories
by
using
small
molecules
recombinant
proteins
to
guide
differentiation
cells
toward
relevant
CNS
phenotypes.
However,
many
these
protocols
fail
recapitulate
cell-guided
programs
intrinsic
embryonic
development,
particularly
signaling
centers
that
emerge
within
neural
tube
during
formation.
Located
on
ventral
end
tube,
floor
plate
acts
as
one
such
center
pattern
dorsal/ventral
axis
secreting
morphogen
Sonic
Hedgehog
(SHH).
Here,
we
present
a
method
synthetic
Notch
(synNotch)
receptor
platform
regulate
SHH
production
subsequent
fate
specification.
We
show
widely
used
configuration
orthogonal
synNotch
ligand
green
fluorescent
protein
(GFP)
mounted
platelet-derived
growth
factor
receptor-β
transmembrane
chassis
does
not
allow
robust
artificial
synNotch-hPSCs
("receivers")
cocultured
with
ligand-presenting
hPSCs
("senders").
discovered
refined
designs
membrane-bound
GFP-ligand
efficient
activation
hPSC
receivers.
A
variant
this
enhanced
drives
sender:hPSC
receiver
cocultures
gives
rise
plate-like
types
seen
development.
This
revised
potential
morphogenesis
studies
designed
uncover
key
paradigms
human
Abstract
Morphogens,
locally
produced
signaling
molecules,
form
a
concentration
gradient
to
guide
tissue
patterning.
Tissue
patterns
emerge
as
collaboration
between
morphogen
diffusion
and
responsive
cell
behaviors,
but
the
mechanisms
through
which
diffusing
morphogens
define
precise
spatial
amidst
biological
fluctuations
remain
unclear.
To
investigate
how
cells
respond
proteins
generate
patterns,
we
develop
SYMPLE3D,
3D
culture
platform.
By
engineering
gene
expression
artificial
morphogens,
observe
that
coupling
signals
with
cadherin-based
adhesion
is
sufficient
convert
into
distinct
domains.
Morphogen-induced
cadherins
gather
activated
single
domain,
removing
ectopically
cells.
In
addition,
reveal
switch-like
induction
of
cadherin-mediated
compaction
mixing,
homogenizing
within
uniformly
domain
sharp
boundary.
These
findings
highlight
cooperation
gradients
in
robust
patterning
introduce
novel
method
for
new
domains
organoids.
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2024,
Номер
unknown
Опубликована: Март 12, 2024
Abstract
Synthetic
developmental
biology
uses
engineering
approaches
to
understand
multicellularity
with
goals
ranging
from
recapitulating
development
building
synthetic
organisms.
Current
include
multicellular
patterning,
controlling
differentiation,
and
implementing
cooperative
cellular
behaviors
in
model
systems.
tools
enable
these
pursuits
genetic
circuits
that
drive
customized
responses
arbitrary
stimuli,
receptors
orthogonal
signaling
channels,
light-
or
drug-inducible
systems
precise
spatial
temporal
control
of
cell
function.
Mouse
embryonic
stem
cells
(mESCs)
offer
a
well-studied
genetically
tractable
pluripotent
chassis
for
pursuing
questions
however,
there
is
minimal
characterization
existing
mESCs
we
lack
toolkits
rapid
iterative
workflows.
Here,
began
address
this
challenge
by
characterizing
small
molecule
contact-inducible
gene
expression
differentiation
mESCs.
We
show
cell-contact
inducible
work
reliably
efficiently
payloads.
Furthermore,
can
direct
into
neurons.
Each
readily
be
used
on
their
own
combination,
opening
many
possibilities
studying
principles
high
precision.
