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.
ACS Sensors,
Год журнала:
2024,
Номер
9(7), С. 3616 - 3624
Опубликована: Июль 9, 2024
Clustered
Regularly
Interspaced
Short
Palindromic
Repeats-CRISPR-Associated
Protein
(CRISPR-Cas)
systems
have
evolved
several
mechanisms
to
specifically
target
foreign
DNA.
These
properties
made
them
attractive
as
biosensors.
The
primary
drawback
associated
with
contemporary
CRISPR-Cas
biosensors
is
their
weak
signaling
capacity,
which
typically
compensated
for
by
coupling
the
nucleic
acid
amplification.
An
alternative
strategy
improve
capacity
engineer
reporter,
i.e.,
design
new
signal-generating
substrates
Cas
proteins.
Unfortunately,
due
reliance
on
custom
synthesis,
most
of
these
engineered
reporter
are
inaccessible
many
researchers.
Herein,
we
investigate
a
substrate
based
fluorescein
(FAM)–tetramethylrhodamine
(TAMRA)
Förster
resonant
energy-transfer
(FRET)
pair
that
functions
seamless
"drop-in"
replacement
existing
reporters,
without
need
change
any
other
aspect
CRISPR-Cas12a-based
assay.
readily
available
and
employs
FRET
produce
two
signals
upon
cleavage
Cas12a.
use
both
in
ratiometric
manner
provides
improved
assay
performance
decreased
time-to-result
CRISPR-Cas12a
assays
when
compared
traditional
FAM–Black
Hole
Quencher
(BHQ)
quench-based
reporter.
We
comprehensively
characterize
this
better
understand
reasons
benchmark
it
against
current
standard
Finally,
showcase
real-world
utility
employ
Recombinase
Polymerase
Amplification
(RPA)–CRISPR-Cas12a
DNA
Endonuclease-Targeted
CRISPR
Trans
Reporter
(DETECTR)
detect
Human
papillomavirus
patient-derived
samples.
ACS Synthetic Biology,
Год журнала:
2024,
Номер
13(6), С. 1633 - 1646
Опубликована: Июнь 11, 2024
A
growing
number
of
applications
require
simultaneous
detection
multiplexed
nucleic
acid
targets
in
a
single
reaction,
which
enables
higher
information
density
combination
with
reduced
assay
time
and
cost.
Clustered
regularly
interspaced
short
palindromic
repeats
(CRISPR)
the
CRISPR-Cas
system
have
broad
for
acids
due
to
their
strong
specificity,
high
sensitivity,
excellent
programmability.
However,
realizing
is
still
challenging
nonspecific
collateral
cleavage
activity,
limited
signal
reporting
strategies,
possible
cross-reactions.
In
this
review,
we
summarize
principles,
features
based
on
further
discuss
challenges
perspective.
Analytical Methods,
Год журнала:
2024,
Номер
unknown
Опубликована: Янв. 1, 2024
Reviews
the
development
of
visual
detection
methods
in
recent
years
based
on
different
Cas
and
analyzes
their
advantages
disadvantages
as
well
challenges
future
research.
Analytical Chemistry,
Год журнала:
2024,
Номер
96(40), С. 16017 - 16026
Опубликована: Сен. 26, 2024
Diagnostic
methods
based
on
CRISPR
technology
have
shown
great
potential
due
to
their
highly
specific,
efficient,
and
sensitive
detection
capabilities.
Although
the
majority
of
current
studies
rely
fluorescent
dye-quencher
reporters,
limitations
dyes,
such
as
poor
photostability
small
Stokes
shifts,
urgently
necessitate
optimization
reporters.
In
this
study,
we
developed
innovative
quantum
dot
(QD)
reporters
for
CRISPR/Cas
systems,
which
not
only
leveraged
advantages
high
photoluminescence
yield
large
shifts
QDs
but
were
also
easily
synthesized
through
a
simple
one-step
hydrothermal
method.
Based
trans-cleavage
characteristics
Cas12a
Cas13a,
two
types
QD
designed,
short
DNA
strand
hybridization-based
achieving
RNA
at
pM
level,
respectively,
validating
performance
in
analysis
clinical
samples.
Furthermore,
unique
property
that
allowed
multicolor
emission
under
one
excitation,
application
simultaneous
diseases
was
further
investigated.
Taken
together,
work
proposed
novel
could
be
applied
various
providing
new
toolbox
expand
diagnosis
bioanalytical
biomedical
fields.
Bioconjugate Chemistry,
Год журнала:
2025,
Номер
unknown
Опубликована: Янв. 21, 2025
Erythropoietin
(EPO)-induced
cellular
signaling
through
the
EPO
receptor
(EPOR)
is
a
fundamental
pathway
for
modulation
of
behavior
and
activity.
In
our
previous
work,
we
showed
in
primary
human
astrocytes
that
multivalent
display
on
surface
semiconductor
quantum
dots
(QDs)
mediates
augmented
JAK/STAT
signaling,
concomitant
1.8-fold
increase
expression
aquaporin-4
(AQPN-4)
channel
proteins,
2-fold
AQPN-4-mediated
water
transport
Our
hypothesis
this
enhanced
involves
simultaneous
ligation
clustering
EPOR
by
QD-EPO
conjugates.
Here,
utilized
embryonic
kidney
(HEK
293T/17)
cell
line
transfected
with
fused
to
green
fluorescent
protein
(eGFP)
visualize
clustering.
We
demonstrate
QDs
displaying
five
copies
(bearing
C-terminal
6-histidine
tract)
nanoparticle
induce
compared
monomeric
at
same
concentration.
findings
confirm
critical
role
played
mediating
EPOR.
More
generally,
these
results
illustrate
capability
nanoparticle-growth
factor
bioconjugates
control
activity
cognate
receptors
important
selective
delivery
signaling.
Smartphones
have
the
potential
to
transform
chemical
and
biological
analysis
by
functioning
as
portable,
versatile,
democratized
platforms
for
molecular
diagnostics.
Recent
advances
in
optical
detection
devices
are
reviewed.
