Nature Biomedical Engineering,
Journal Year:
2024,
Volume and Issue:
unknown
Published: July 10, 2024
Abstract
Prime
editing
(PE)
enables
precise
and
versatile
genome
without
requiring
double-stranded
DNA
breaks.
Here
we
describe
the
systematic
optimization
of
PE
systems
to
efficiently
correct
human
cystic
fibrosis
(CF)
transmembrane
conductance
regulator
(
CFTR
)
F508del,
a
three-nucleotide
deletion
that
is
predominant
cause
CF.
By
combining
six
efficiency
optimizations
for
PE—engineered
guide
RNAs,
PEmax
architecture,
transient
expression
dominant-negative
mismatch
repair
protein,
strategic
silent
edits,
PE6
variants
proximal
‘dead’
single-guide
RNAs—we
increased
correction
efficiencies
F508del
from
less
than
0.5%
in
HEK293T
cells
58%
immortalized
bronchial
epithelial
(a
140-fold
improvement)
25%
patient-derived
airway
cells.
The
also
resulted
minimal
off-target
editing,
edit-to-indel
ratios
3.5-fold
greater
those
achieved
by
nuclease-mediated
homology-directed
repair,
functional
restoration
ion
channels
over
50%
wild-type
levels
(similar
via
combination
treatment
with
elexacaftor,
tezacaftor
ivacaftor)
primary
Our
findings
support
feasibility
durable
one-time
Signal Transduction and Targeted Therapy,
Journal Year:
2024,
Volume and Issue:
9(1)
Published: Feb. 21, 2024
Abstract
Inflammation-associated
diseases
encompass
a
range
of
infectious
and
non-infectious
inflammatory
diseases,
which
continuously
pose
one
the
most
serious
threats
to
human
health,
attributed
factors
such
as
emergence
new
pathogens,
increasing
drug
resistance,
changes
in
living
environments
lifestyles,
aging
population.
Despite
rapid
advancements
mechanistic
research
development
for
these
current
treatments
often
have
limited
efficacy
notable
side
effects,
necessitating
more
effective
targeted
anti-inflammatory
therapies.
In
recent
years,
nanotechnology
has
provided
crucial
technological
support
prevention,
treatment,
detection
inflammation-associated
diseases.
Various
types
nanoparticles
(NPs)
play
significant
roles,
serving
vaccine
vehicles
enhance
immunogenicity
carriers
improve
targeting
bioavailability.
NPs
can
also
directly
combat
pathogens
inflammation.
addition,
facilitated
biosensors
pathogen
imaging
techniques
This
review
categorizes
characterizes
different
NPs,
summarizes
their
applications
It
discusses
challenges
associated
with
clinical
translation
this
field
explores
latest
developments
prospects.
conclusion,
opens
up
possibilities
comprehensive
management
Cell,
Journal Year:
2024,
Volume and Issue:
187(5), P. 1076 - 1100
Published: Feb. 1, 2024
Genome
editing
has
been
a
transformative
force
in
the
life
sciences
and
human
medicine,
offering
unprecedented
opportunities
to
dissect
complex
biological
processes
treat
underlying
causes
of
many
genetic
diseases.
CRISPR-based
technologies,
with
their
remarkable
efficiency
easy
programmability,
stand
at
forefront
this
revolution.
In
Review,
we
discuss
current
state
CRISPR
gene
technologies
both
research
therapy,
highlighting
limitations
that
constrain
them
technological
innovations
have
developed
recent
years
address
them.
Additionally,
examine
summarize
landscape
applications
context
health
therapeutics.
Finally,
outline
potential
future
developments
could
shape
coming
years.
Cell,
Journal Year:
2023,
Volume and Issue:
186(18), P. 3983 - 4002.e26
Published: Aug. 1, 2023
Prime
editing
enables
a
wide
variety
of
precise
genome
edits
in
living
cells.
Here
we
use
protein
evolution
and
engineering
to
generate
prime
editors
with
reduced
size
improved
efficiency.
Using
phage-assisted
evolution,
efficiencies
compact
reverse
transcriptases
by
up
22-fold
generated
that
are
516–810
base
pairs
smaller
than
the
current-generation
editor
PEmax.
We
discovered
different
specialize
types
used
this
insight
outperform
PEmax
PEmaxΔRNaseH,
truncated
dual-AAV
delivery
systems.
Finally,
Cas9
domains
improve
editing.
These
resulting
(PE6a-g)
enhance
therapeutically
relevant
patient-derived
fibroblasts
primary
human
T-cells.
PE6
variants
also
enable
longer
insertions
be
installed
vivo
following
delivery,
achieving
40%
loxP
insertion
cortex
murine
brain,
24-fold
improvement
compared
previous
state-of-the-art
editors.
Nature Biotechnology,
Journal Year:
2023,
Volume and Issue:
42(2), P. 253 - 264
Published: May 4, 2023
Abstract
Realizing
the
promise
of
prime
editing
for
study
and
treatment
genetic
disorders
requires
efficient
methods
delivering
editors
(PEs)
in
vivo.
Here
we
describe
identification
bottlenecks
limiting
adeno-associated
virus
(AAV)-mediated
vivo
development
AAV-PE
vectors
with
increased
PE
expression,
guide
RNA
stability
modulation
DNA
repair.
The
resulting
dual-AAV
systems,
v1em
v3em
PE-AAV,
enable
therapeutically
relevant
mouse
brain
(up
to
42%
efficiency
cortex),
liver
46%)
heart
11%).
We
apply
these
systems
install
putative
protective
mutations
Alzheimer’s
disease
astrocytes
coronary
artery
hepatocytes.
In
PE-AAV
caused
no
detectable
off-target
effects
or
significant
changes
enzymes
histology.
Optimized
support
highest
unenriched
levels
reported
date,
facilitating
potential
diseases
a
component.
Nature Communications,
Journal Year:
2023,
Volume and Issue:
14(1)
Published: Jan. 13, 2023
CRISPR-Cas
gene
editing
has
revolutionized
experimental
molecular
biology
over
the
past
decade
and
holds
great
promise
for
treatment
of
human
genetic
diseases.
Here
we
review
development
CRISPR-Cas9/Cas12/Cas13
nucleases,
DNA
base
editors,
prime
RNA
focusing
on
assessment
improvement
their
precision
safety,
pushing
limit
specificity
efficiency.
We
summarize
capabilities
limitations
each
CRISPR
tool
from
to
editing,
highlight
opportunities
future
improvements
applications
in
basic
research,
as
well
therapeutic
clinical
considerations
use
patients.
