bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2024,
Номер
unknown
Опубликована: Авг. 12, 2024
ABSTRACT
Cystic
fibrosis
(CF)
is
caused
by
mutations
in
the
cystic
transmembrane
conductance
regulator
(
CFTR
)
gene.
Although
many
people
with
CF
(pwCF)
are
treated
using
modulators,
some
non-responsive
due
to
their
genotype
or
other
uncharacterized
reasons.
Autologous
airway
stem
cell
therapies,
which
cDNA
has
been
replaced,
may
enable
a
durable
therapy
for
all
pwCF.
Previously,
CRISPR-Cas9
two
AAVs
was
used
sequentially
insert
halves
of
and
an
enrichment
cassette
into
locus.
However,
editing
efficiency
<10%
required
restore
function.
Further
improvement
gene
insertion
enhance
production.
To
improve
human
basal
cells
(ABCs),
we
evaluated
use
small
molecules
AZD7648
ART558
inhibit
non-homologous
end
joining
(NHEJ)
micro-homology
mediated
(MMEJ).
Adding
alone
improved
2-3-fold.
both
but
induced
toxicity.
ABCs
edited
presence
produced
differentiated
epithelial
sheets
restored
function
after
enrichment.
did
not
increase
off-target
editing.
studies
necessary
validate
if
treatment
enriches
oncogenic
mutations.
In
the
past
10
years,
CRISPR-Cas9
has
revolutionized
gene-editing
field
due
to
its
modularity,
simplicity,
and
efficacy.
It
been
applied
for
creation
of
in
vivo
models,
further
understand
human
biology,
toward
curing
genetic
diseases.
However,
there
remain
significant
delivery
barriers
application
clinic,
especially
extrahepatic
applications.
this
work,
high-throughput
molecular
barcoding
techniques
were
used
alongside
traditional
screening
methodologies
simultaneously
evaluate
LNP
formulations
encapsulating
ribonucleoproteins
(RNPs)
vitro
efficiency
biodistribution.
This
resulted
identification
a
lung-tropic
formulation,
which
shows
efficient
gene
editing
endothelial
epithelial
cells
within
lung,
targeting
both
model
reporter
clinically
relevant
genomic
targets.
Further,
no
off-target
indel
formation
liver,
making
it
highly
specific
system
lung-editing
Pharmaceutics,
Год журнала:
2025,
Номер
17(1), С. 104 - 104
Опубликована: Янв. 14, 2025
Monogenic
disorders
are
a
group
of
human
diseases
caused
by
mutations
in
single
genes.
While
some
disease-altering
treatments
offer
relief
and
slow
the
progression
certain
conditions,
majority
monogenic
still
lack
effective
therapies.
In
recent
years,
gene
therapy
has
appeared
as
promising
approach
for
addressing
genetic
disorders.
However,
despite
advancements
manipulation
tools
delivery
systems,
several
challenges
remain
unresolved,
including
inefficient
delivery,
sustained
expression,
immunogenicity,
toxicity,
capacity
limitations,
genomic
integration
risks,
limited
tissue
specificity.
This
review
provides
an
overview
plasmid-based
techniques
methods
currently
employed
diseases,
highlighting
they
face
exploring
potential
strategies
to
overcome
these
barriers.
iScience,
Год журнала:
2025,
Номер
28(3), С. 111979 - 111979
Опубликована: Фев. 21, 2025
Cystic
fibrosis
(CF)
is
a
life-shortening
autosomal
recessive
disease,
caused
by
loss-of-function
mutations
that
affect
the
CF
transmembrane
conductance
regulator
(CFTR)
anion
channel.
G542X
second-most
common
CF-causing
variant,
and
it
does
not
respond
to
current
CFTR
modulator
drugs.
Our
study
explores
use
of
adenine
base
editing
edit
non-CF-causing
G542R,
recover
function.
Using
editor
engineered
virus-like
particles
(BE-eVLPs)
in
patient-derived
intestinal
organoids,
we
achieved
∼2%
G542X-to-G542R
efficiency
restored
CFTR-mediated
chloride
transport
∼6.4%
wild-type
levels,
independent
treatment,
with
no
bystander
edits.
This
proof-of-principle
demonstrates
potential
rescue
provides
foundation
for
future
-
vivo
applications.
Abstract
Gut
organoids
are
3D
cellular
structures
derived
from
adult
or
pluripotent
stem
cells,
capable
of
closely
replicating
the
physiological
properties
gut.
These
serve
as
powerful
tools
for
studying
gut
development
and
modeling
pathogenesis
intestinal
diseases.
This
review
provides
an
in-depth
exploration
technological
advancements
applications
organoids,
with
a
focus
on
their
construction
methods.
Additionally,
potential
in
disease
modeling,
microenvironmental
simulation,
personalized
medicine
summarized.
aims
to
offer
perspectives
directions
understanding
mechanisms
health
well
developing
innovative
therapeutic
strategies.
Cells,
Год журнала:
2025,
Номер
14(6), С. 457 - 457
Опубликована: Март 19, 2025
Colorectal
epithelium
was
the
first
long-term
3D
organoid
culture
established
in
vitro.
Identification
of
key
components
essential
for
survival
stem
cell
niche
allowed
an
indefinite
propagation
these
cultures
and
modulation
their
differentiation
into
various
lineages
mature
intestinal
epithelial
cells.
While
methods
were
eventually
adapted
to
establish
organoids
from
different
organs,
colorectal
remain
a
pioneering
model
development
new
applications
health
disease.
Several
basic
applicative
aspects
culture,
modeling,
monitoring
testing
are
analyzed
this
review.
We
also
tackle
ethical
problems
biobanking
distribution
precious
research
tools,
frequently
confined
laboratory
origin
or
condemned
destruction
at
end
project.
