Degron tagging for rapid protein degradation in mice
Disease Models & Mechanisms,
Год журнала:
2024,
Номер
17(4)
Опубликована: Апрель 1, 2024
Degron
tagging
allows
proteins
of
interest
to
be
rapidly
degraded,
in
a
reversible
and
tuneable
manner,
response
chemical
stimulus.
This
provides
numerous
opportunities
for
understanding
disease
mechanisms,
modelling
therapeutic
interventions
constructing
synthetic
gene
networks.
In
recent
years,
many
laboratories
have
applied
degron
successfully
cultured
mammalian
cells,
spurred
by
rapid
advances
the
fields
genome
editing
targeted
protein
degradation.
this
At
Glance
article,
we
focus
on
efforts
apply
mouse
models,
discussing
distinct
set
challenges
posed
vivo
environment.
Язык: Английский
PROTAC-Based Protein Degradation as a Promising Strategy for Targeted Therapy in Sarcomas
International Journal of Molecular Sciences,
Год журнала:
2023,
Номер
24(22), С. 16346 - 16346
Опубликована: Ноя. 15, 2023
Sarcomas
are
heterogeneous
bone
and
soft
tissue
cancers
representing
the
second
most
common
tumor
type
in
children
adolescents.
Histology
genetic
profiling
discovered
more
than
100
subtypes,
which
characterized
by
peculiar
molecular
vulnerabilities.
However,
limited
therapeutic
options
exist
beyond
standard
therapy
clinical
benefits
from
targeted
therapies
were
observed
only
a
minority
of
patients
with
sarcomas.
The
rarity
these
tumors,
paucity
actionable
mutations,
limitations
chemical
composition
current
hindered
use
approaches
Targeted
protein
degradation
(TPD)
is
an
innovative
pharmacological
modality
to
directly
alter
abundance
promising
potential
cancer,
even
for
undruggable
proteins.
TPD
based
on
small
molecules
called
degraders
or
proteolysis-targeting
chimeras
(PROTACs),
trigger
ubiquitin-dependent
interest.
In
this
review,
we
will
discuss
major
features
PROTAC
PROTAC-derived
systems
target
validation
cancer
treatment
focus
overcome
issues
connected
sarcomas,
including
drug
resistance,
specificity,
targets.
A
deeper
understanding
strategies
might
provide
new
fuel
drive
personalized
medicine
Язык: Английский
Differential Protein Degradation with a Super-Degron Tag and Thalidomide Analogs: EGFP is Degraded in Wild-Type Mouse Cell, While PD-1 Requires CRBN Humanization, Providing New Insights into T Cell Regulation
Опубликована: Янв. 1, 2025
It
was
previously
considered
that
protein
knockdown
using
a
combination
of
zinc
finger
degron
tag
and
thalidomide
analog
is
impossible
in
mouse
cells,
however,
EGFP
expression
as
an
indicator,
we
found
possible
super-degron
(SD)
with
iberdomide
or
mezigdomide
cells.
Despite
the
efficient
degradation
SD-tagged
wild-type
PD-1
could
not
be
degraded
but
human
Jurkat
cells
human-type
CEREBLON
(CRBNI391V).
In
mice
CRBNI391V,
endogenous
tagged
SD
efficiently
suppressed
T
both
cultured
whole
body.
addition,
comparison
treatment
anti-PD-1
antibody
suggested
transient
downregulation
might
prevent
cell
exhaustion
compared
to
treatment.
brain
also
reduced
by
pomalidomide
crosses
brain-blood
barrier.
These
results
suggest
analogs
are
effective
for
vitro
vivo
knockdown,
although
some
conditional
settings
required.
Язык: Английский
Rapid and sustained degradation of the essential centrosome protein CEP192 in live mice using the AID2 system
Science Advances,
Год журнала:
2025,
Номер
11(9)
Опубликована: Фев. 28, 2025
Studying
essential
genes
required
for
dynamic
processes
in
live
mice
is
challenging
as
genetic
perturbations
are
irreversible
and
limited
by
slow
protein
depletion
kinetics.
The
auxin-inducible
degron
(AID)
system
a
powerful
tool
analyzing
inducible
loss
vitro,
but
it
toxic
to
mice.
Here,
we
use
an
optimized
second-generation
AID
achieve
the
conditional
reversible
of
centrosomal
CEP192
We
show
that
auxin
derivative
5-phenyl-indole-3-acetic
acid
well
tolerated
over
2
weeks
drives
near-complete
degradation
less
than
1
hour
vivo.
did
not
affect
centriole
duplication
decreased
γ-tubulin
recruitment
centrosomes
impairing
mitotic
spindle
assembly.
Sustained
vivo
led
cell
division
failure
death
proliferative
tissues.
Thus,
suited
rapid
and/or
sustained
study
functions
Язык: Английский
Drug discovery technologies–Current and future trends
Elsevier eBooks,
Год журнала:
2025,
Номер
unknown
Опубликована: Янв. 1, 2025
Язык: Английский
Establishment and characterization of mouse lines useful for endogenous protein degradation via an improved auxin‐inducible degron system (AID2)
Hatsune Makino‐Itou,
Noriko Yamatani,
Akemi Okubo
и другие.
Development Growth & Differentiation,
Год журнала:
2024,
Номер
66(7), С. 384 - 393
Опубликована: Сен. 1, 2024
Abstract
The
development
of
new
technologies
opens
avenues
in
the
research
field.
Gene
knockout
is
a
key
method
for
analyzing
gene
function
mice.
Currently,
conditional
strategies
are
employed
to
examine
temporal
and
spatial
function.
However,
phenotypes
sometimes
not
observed
because
time
required
depletion
due
long
half‐life
target
proteins.
Protein
knockdown
using
an
improved
auxin‐inducible
degron
system,
AID2,
overcomes
such
difficulties
owing
rapid
efficient
depletion.
We
AID‐tagged
proteins
within
few
several
hours
by
simple
intraperitoneal
injection
auxin
analog,
5‐Ph‐IAA,
which
much
shorter
than
knockout.
Importantly,
loss
protein
reversible,
making
useful
measure
effects
transient
Here,
we
also
established
mouse
lines
AID2‐medicated
knockdown,
include
knock‐in
ROSA26
locus;
one
expresses
TIR1(F74G),
other
reporter
expressing
AID‐mCherry.
germ‐cell‐specific
TIR1
line
confirmed
specificity.
In
addition,
introduced
AID
tag
endogenous
protein,
DCP2
via
CAS9‐mediated
editing
method.
that
was
effectively
eliminated
resulted
similar
phenotype
20
h.
Язык: Английский
The AID2 system offers a potent tool for rapid, reversible, or sustained degradation of essential proteins in live mice
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2024,
Номер
unknown
Опубликована: Июнь 5, 2024
Abstract
Studying
essential
genes
required
for
dynamic
processes
in
live
mice
is
challenging
as
genetic
perturbations
are
irreversible
and
limited
by
slow
protein
depletion
kinetics.
The
first-generation
auxin-inducible-degron
(AID)
system
a
powerful
tool
analyzing
inducible
loss
cultured
cells.
However,
auxin
administration
toxic
to
mice,
preventing
its
long-term
use
animals.
Here,
we
an
optimized
second-generation
AID
achieve
the
conditional
reversible
of
centrosomal
CEP192
mice.
We
show
that
derivative
5-Ph-IAA
well
tolerated
over
two
weeks
drives
near-complete
CEP192-mAID
degradation
less
than
one
hour
vivo
.
Prolonged
led
cell
division
failure
death
proliferative
tissues.
Thus,
suited
rapid
and/or
sustained
offering
valuable
new
interrogating
function
Язык: Английский