Analytical Chemistry,
Journal Year:
2025,
Volume and Issue:
unknown
Published: March 31, 2025
Viral
proteases
are
critical
molecular
targets
in
viral
pathogenesis,
representing
pivotal
biomarkers
for
understanding
infection
mechanisms
and
developing
antiviral
therapeutics.
This
study
introduces
a
label-free
electrochemical
biosensor
that
enables
sensitive
protease
detection
by
integrating
protease-responsive
CRISPR/Cas
protein
switches
(CasPSs)
with
hemin
aptamer-functionalized
interface.
The
biosensor's
mechanism
relies
on
protease-mediated
proteolysis,
which
leads
to
the
release
of
active
Cas12a
proteins
from
CasPSs
generates
amplified
responses
through
continuous
cleavage
immobilized
redox-active
hemin/aptamer
complexes.
achieved
specific
hepatitis
C
virus
NS3/4A
sensing
femtomolar
sensitivity
could
be
readily
expanded
other
replacing
CasPS
module.
feasibility
this
was
demonstrated
monitoring
enterovirus
71
3C
activities
virus-infected
cell
samples
different
loads
postinfection
times.
provides
promising
strategy
CRISPR
biosensing
platforms,
offering
helpful
analytical
tool
drug
screening.
Angewandte Chemie International Edition,
Journal Year:
2024,
Volume and Issue:
63(16)
Published: Feb. 26, 2024
Abstract
Proteins
capable
of
switching
between
distinct
active
states
in
response
to
biochemical
cues
are
ideal
for
sensing
and
controlling
biological
processes.
Activatable
CRISPR‐Cas
systems
significant
precise
genetic
manipulation
sensitive
molecular
diagnostics,
yet
directly
Cas
protein
function
remains
challenging.
Herein,
we
explore
anti‐CRISPR
(Acr)
proteins
as
modules
create
synthetic
switches
(CasPSs)
based
on
computational
chemistry‐directed
rational
interface
engineering.
Guided
by
fingerprint
analysis,
electrostatic
potential
mapping,
binding
free
energy
calculations,
rationally
engineer
the
interaction
Cas12a
its
cognate
Acr
(AcrVA4
AcrVA5)
generate
a
series
orthogonal
protease‐responsive
CasPSs.
These
CasPSs
enable
conversion
specific
proteolytic
events
into
activation
with
high
ratios
(up
34.3‐fold).
advancements
proteolysis‐inducible
genome
editing
mammalian
cells
detection
viral
protease
activities
during
virus
infection.
This
work
provides
promising
strategy
developing
tools
controllable
gene
regulation
clinical
diagnostics.
Journal of Agricultural and Food Chemistry,
Journal Year:
2024,
Volume and Issue:
72(14), P. 8257 - 8268
Published: March 26, 2024
Rapid
and
accurate
detection
of
the
zoonotic
nematode
Anisakis
is
poised
to
control
its
epidemic.
The
clustered
regularly
interspaced
short
palindromic
repeats
(CRISPR)/Cas-associated
assay
shows
great
potential
in
pathogenic
microorganisms.
one-tube
method
integrated
CRISPR
system
with
recombinase
polymerase
amplification
(RPA)
avoid
risk
aerosol
pollution;
however,
it
suffers
from
low
sensitivity
due
incompatibility
two
systems
additional
manual
operations.
Therefore,
present
study,
agarose
hydrogel
boosted
RPA-CRISPR/Cas12a
was
constructed
by
adding
hydrogel,
which
avoided
initially
efficiency
RPA
caused
cleavage
Cas12a
achieved
reaction
continuity.
10-fold
higher
than
that
system.
This
used
for
within
80
min
sample
result,
achieving
point-of-care
testing
(POCT)
through
a
smartphone
portable
device.
study
provided
novel
toolbox
POCT
significant
application
value
preventing
infection.
Viruses,
Journal Year:
2024,
Volume and Issue:
16(5), P. 753 - 753
Published: May 10, 2024
Avian
influenza
viruses
(AIVs)
of
the
H5
subtype
rank
among
most
serious
pathogens,
leading
to
significant
economic
losses
in
global
poultry
industry
and
posing
risks
human
health.
Therefore,
rapid
accurate
virus
detection
is
crucial
for
prevention
control
AIVs.
In
this
study,
we
established
a
novel
method
by
utilizing
precision
CRISPR/Cas12a
efficiency
RT-RPA
technologies.
This
assay
facilitates
direct
visualization
results
through
blue
light
lateral
flow
strips,
accurately
identifying
with
high
specificity
without
cross-reactivity
against
other
AIV
subtypes,
NDV,
IBV,
IBDV.
With
thresholds
1.9
copies/μL
(blue
light)
×
103
(lateral
strips),
our
not
only
competes
but
also
slightly
surpasses
RT-qPCR,
demonstrating
an
80.70%
positive
rate
across
81
clinical
samples.
The
RT-RPA/CRISPR-based
characterized
sensitivity,
specificity,
independence
from
specialized
equipment.
immediate
field
applicability
RT-RPA/CRISPR
approach
underscores
its
importance
as
effective
tool
early
management
outbreaks
caused
