Nanomaterials-integrated
CRISPR/Cas
systems
have
rapidly
emerged
as
powerful
next-generation
platforms
for
optical
biosensing.
These
integrated
harness
the
precision
of
CRISPR/Cas-mediated
nucleic
acid
detection
while
leveraging
unique
properties
nanomaterials
to
achieve
enhanced
sensitivity
and
expanded
analytical
capabilities,
thereby
broadening
their
diagnostic
potential.
By
incorporating
a
diverse
range
nanomaterials,
these
effectively
expand
toolbox
detection,
offering
adaptable
solutions
tailored
various
challenges.
This
review
provides
comprehensive
overview
successfully
into
CRISPR/Cas-based
sensing
systems.
It
examines
multiple
modalities,
including
fluorescence,
electrochemiluminescence,
colorimetry,
surface-enhanced
Raman
spectroscopy,
highlighting
how
facilitate
signal
amplification,
enable
multiplexing,
support
development
point-of-care
applications.
Additionally,
practical
applications
in
critical
fields
such
healthcare
diagnostics
environmental
monitoring
are
showcased.
While
offer
considerable
advantages,
several
real-world
challenges
complexity
assay
workflows,
impact
cost,
regulatory
hurdles
must
be
addressed
before
widespread
implementation
can
achieved.
identifying
obstacles
proposing
strategic
solutions,
we
aim
pave
way
continued
advancement
adoption
nanomaterial-integrated
biosensing
technologies.
Analytical Chemistry,
Год журнала:
2024,
Номер
unknown
Опубликована: Дек. 5, 2024
Circulating
tumor
nucleic
acids
(CTNAs),
which
consist
of
cell-free
DNA
or
RNA
released
from
cells,
are
utilized
as
potential
biomarkers
for
diagnosing
and
managing
prognosis.
There
is
a
significant
demand
developing
highly
sensitive
reliable
assay
CTNAs
detection.
In
this
study,
we
engineered
CRISPR/Cas12a
corona
nanomachine
capable
detecting
circulating
in
serum.
This
consists
protein
shell
incorporating
Cas12a/crRNA
ribonucleoprotein
complexes
scaffold
AuNP
core
decorated
with
substrate
ssDNA
strands.
The
protective
CRISPR
shields
the
acid
degradation
by
nuclease
DNase/RNase,
thereby
enhancing
stability
biological
fluids,
even
tolerating
up
to
undiluted
human
serum
FBS.
Upon
encountering
target
CTNAs,
activated
through
sequence-specific
hybridization
between
crRNA
CTNAs.
Subsequently,
autonomously
cleaves
collateral
substrates
on
AuNPs,
releasing
fluorophore-labeled
fragment
generating
an
increasing
fluorescent
signal.
was
successfully
employed
detect
various
including
(ct)DNA/RNA
(EGFR
L858R)
microRNA-21,
achieving
limit
detection
0.14
pM
ctDNA
1.0
RNA.
enables
simultaneous
both
complex
samples,
offering
promising
tool
early
diagnosis.
Analytical Chemistry,
Год журнала:
2025,
Номер
unknown
Опубликована: Март 11, 2025
CRISPR-Cas
systems
represent
a
highly
programmable
and
precise
nucleic
acid-targeting
platform,
which
has
been
strategically
engineered
as
versatile
toolkit
for
biosensing
bioimaging
applications.
Nevertheless,
their
analytical
performance
is
constrained
by
inherent
functional
activity
limitations
of
natural
CRISPR/Cas
systems,
underscoring
the
critical
role
molecular
engineering
in
enhancing
capabilities.
This
review
comprehensively
examines
recent
advancements
ribonucleoproteins
(RNPs)
to
enhance
capabilities
advanced
detection
cellular
imaging.
We
explore
innovative
strategies
developing
enhanced
RNPs,
including
Cas
protein
through
mutagenesis
fusion
techniques,
guide
RNA
via
chemical
structural
modifications.
Furthermore,
we
evaluate
these
RNPs'
applications
sensitive
biomarker
live-cell
genomic
DNA
monitoring,
while
analyzing
current
challenges
prospective
developments
RNP
bioimaging.
Analytical Chemistry,
Год журнала:
2025,
Номер
unknown
Опубликована: Март 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.
Nanomaterials-integrated
CRISPR/Cas
systems
have
rapidly
emerged
as
powerful
next-generation
platforms
for
optical
biosensing.
These
integrated
harness
the
precision
of
CRISPR/Cas-mediated
nucleic
acid
detection
while
leveraging
unique
properties
nanomaterials
to
achieve
enhanced
sensitivity
and
expanded
analytical
capabilities,
thereby
broadening
their
diagnostic
potential.
By
incorporating
a
diverse
range
nanomaterials,
these
effectively
expand
toolbox
detection,
offering
adaptable
solutions
tailored
various
challenges.
This
review
provides
comprehensive
overview
successfully
into
CRISPR/Cas-based
sensing
systems.
It
examines
multiple
modalities,
including
fluorescence,
electrochemiluminescence,
colorimetry,
surface-enhanced
Raman
spectroscopy,
highlighting
how
facilitate
signal
amplification,
enable
multiplexing,
support
development
point-of-care
applications.
Additionally,
practical
applications
in
critical
fields
such
healthcare
diagnostics
environmental
monitoring
are
showcased.
While
offer
considerable
advantages,
several
real-world
challenges
complexity
assay
workflows,
impact
cost,
regulatory
hurdles
must
be
addressed
before
widespread
implementation
can
achieved.
identifying
obstacles
proposing
strategic
solutions,
we
aim
pave
way
continued
advancement
adoption
nanomaterial-integrated
biosensing
technologies.
