Analytical Chemistry,
Год журнала:
2021,
Номер
93(30), С. 10719 - 10726
Опубликована: Июль 19, 2021
A
CRISPR-Cas
system
holds
great
promise
as
a
next-generation
biosensing
technology
for
molecular
diagnostics.
Nevertheless,
the
current
CRISPR-Cas12a-based
detection
strategies
always
need
bulky
instruments
or
auxiliary
devices
to
obtain
quantitative
signal
output,
which
restrains
its
point-of-care
testing
application.
Herein,
we
proposed
duplex-specific
nuclease-assisted
CRISPR-Cas12a
strategy
detect
microRNA
(miRNA)
with
personal
glucose
meter.
The
target
miRNA
was
first
converted
into
an
amplified
initiator
DNA
via
nuclease.
Afterward,
activated
collateral
cleavage
activity
of
cleave
single-strand
(ssDNA)
linker
on
sucrase-ssDNA-modified
magnetic
beads,
led
release
sucrase.
released
sucrase
collected
and
then
utilized
catalyze
sucrose
glucose,
could
be
quantitatively
detected
by
change
in
directly
reflected
concentration
miRNA,
avoided
expensive
equipment
quantification.
Two
different
miRNAs
(miRNA21
miRNA205)
simply
changing
sequence
template
strand
(H
strand).
developed
showed
high
sensitivity
limit
(LOD)
2.4
1.1
pM
miRNA21
miRNA205,
respectively.
In
addition,
good
selectivity
anti-interference
ability
were
achieved
using
this
method,
enabled
it
promising
at
point-of-care.
Nano Letters,
Год журнала:
2024,
Номер
24(7), С. 2360 - 2368
Опубликована: Фев. 12, 2024
Accurate
and
sensitive
analysis
of
circulating
tumor
cells
(CTCs)
in
human
blood
provides
a
non-invasive
approach
for
the
evaluation
cancer
metastasis
early
diagnosis.
Herein,
we
demonstrate
controllable
assembly
quantum
dot
(QD)-based
aptasensor
guided
by
CRISPR/Cas12a
direct
measurement
CTCs
blood.
We
introduce
magnetic
bead@activator/recognizer
duplex
core–shell
structure
to
construct
multifunctional
platform
capture
detection
blood,
without
need
additional
CTC
release
re-identification
steps.
Notably,
introduction
separation
ensures
that
only
target-induced
free
activator
can
initiate
downstream
catalysis,
efficiently
avoiding
undesired
catalysis
triggered
inappropriate
recognition
activator/recognizer
crRNAs.
This
achieves
high
CTC-capture
efficiency
(82.72%)
with
limit
2
mL–1
holding
great
promise
liquid
biopsy
cancers.
Analytical Chemistry,
Год журнала:
2025,
Номер
unknown
Опубликована: Янв. 19, 2025
Precise
identification
and
analysis
of
multiple
protein
biomarkers
on
the
surface
breast
cancer
cell-derived
extracellular
vesicles
(BC-EVs)
are
great
significance
for
noninvasive
diagnosis
subtypes,
but
it
remains
a
major
challenge
owing
to
their
high
heterogeneity
low
abundance.
Herein,
we
established
CRISPR-based
homogeneous
electrochemical
strategy
near-zero
background
ultrasensitive
detection
BC-EVs.
To
realize
high-performance
capture
isolation
BC-EVs,
fluidity-enhanced
magnetic
nanoprobes
were
facilely
prepared.
After
capturing
AND
logic
gate-based
catalytic
hairpin
assembly
(CHA)
trans-cleavage
activity
CRISPR-Cas12a
against
signal
triggered
successively,
generating
significant
signal.
Notably,
as-developed
metal-mediated
could
efficiently
decrease
by
separation,
endowing
method
with
signal-to-noise
ratio.
Consequently,
ingeniously
integrating
DNA
CRISPR-CHA
amplification
dual
in
strategy,
precise
BC-EVs
was
successfully
achieved
through
simultaneous
specific
recognition
markers
surface.
More
importantly,
this
approach
effectively
discriminate
subgroups
clinical
serum
samples,
which
may
provide
opportunities
accurate
prognosis
evaluation
manner.
Development
of
a
multiplexed
and
sensitive
biosensing
platform
is
priority
for
public
health
security.
We
report
micropore
resistance
counting
based
on
polystyrene
microsphere
size-based
encoding
Clostridium
butyricum
Argonaute
(CbAgo)
decoding
ultrasensitive
detection.
Initially,
we
constructed
target
DNA-modified
coding
system
counting.
Subsequently,
the
precise
recognition
cleavage
capabilities
guide
DNA-activated
CbAgo
protein
enable
encoded
system.
Changes
in
concentration
microspheres
are
presented
as
signal
readout.
The
demonstrated
excellent
performance
detection
three
mycotoxins
(with
sensitivity
range
over
4
orders
magnitude
reaching
pg/mL
level)
two
inflammatory
markers
at
pg/mL.
Combining
enzyme
by
with
counting,
developed
highly
tool
wide-ranging
potential
applications
such
clinical
diagnosis,
food
safety
inspection,
environmental
monitoring.
Analytical Chemistry,
Год журнала:
2025,
Номер
unknown
Опубликована: Янв. 29, 2025
Detection
and
imaging
of
dual
miRNAs
based
on
AND
logic
gates
can
improve
the
accuracy
early
diagnosis
disease.
However,
a
single
target
may
lead
to
false
positive.
Hence,
this
work
rationally
integrates
hyperbranched
rolling
circle
amplification
(HRCA)
with
Cas12a
by
replacing
PAM
sequence
bubble
sensitively
detect
image
miRNA-10b
miRNA-21
gate.
When
are
both
present,
two
padlocks
linked
into
circular
DNA
as
template
for
RCA.
Long
ssDNA
products
generated
under
catalysis
phi29
polymerase,
which
cis-cleaved
activated
trans-cleavage
generate
fluorescent
signals.
Subsequently,
primer
hybridizes
cis-cleavage
is
extended
dsDNA
substrate
produce
more
produces
significant
signals
leading
positive
due
presence
protospacer
adjacent
motif
(PAM)
padlock.
After
removed
from
padlock,
RCA
form
bubbles
replace
PAM,
activate
without
affecting
sensitivity
reduce
The
introduction
enables
second
utilization
Cas12a,
increasing
signal-to-noise
ratio.
HRCA
exhibit
optimal
activity
in
T4
ligase
buffer,
achieving
one-pot
detection
miRNAs.
In
addition,
HRCA-Cas12a
method
intracellular
visualization
It
exhibits
ability
distinguish
different
types
cancer
cells
expression
level
Analytical Chemistry,
Год журнала:
2021,
Номер
93(30), С. 10719 - 10726
Опубликована: Июль 19, 2021
A
CRISPR-Cas
system
holds
great
promise
as
a
next-generation
biosensing
technology
for
molecular
diagnostics.
Nevertheless,
the
current
CRISPR-Cas12a-based
detection
strategies
always
need
bulky
instruments
or
auxiliary
devices
to
obtain
quantitative
signal
output,
which
restrains
its
point-of-care
testing
application.
Herein,
we
proposed
duplex-specific
nuclease-assisted
CRISPR-Cas12a
strategy
detect
microRNA
(miRNA)
with
personal
glucose
meter.
The
target
miRNA
was
first
converted
into
an
amplified
initiator
DNA
via
nuclease.
Afterward,
activated
collateral
cleavage
activity
of
cleave
single-strand
(ssDNA)
linker
on
sucrase-ssDNA-modified
magnetic
beads,
led
release
sucrase.
released
sucrase
collected
and
then
utilized
catalyze
sucrose
glucose,
could
be
quantitatively
detected
by
change
in
directly
reflected
concentration
miRNA,
avoided
expensive
equipment
quantification.
Two
different
miRNAs
(miRNA21
miRNA205)
simply
changing
sequence
template
strand
(H
strand).
developed
showed
high
sensitivity
limit
(LOD)
2.4
1.1
pM
miRNA21
miRNA205,
respectively.
In
addition,
good
selectivity
anti-interference
ability
were
achieved
using
this
method,
enabled
it
promising
at
point-of-care.