ACS Sensors,
Год журнала:
2023,
Номер
8(10), С. 3623 - 3642
Опубликована: Окт. 11, 2023
Over
the
past
few
decades,
pathogens
have
posed
a
threat
to
human
security,
and
rapid
identification
of
should
be
one
ideal
methods
prevent
major
public
health
security
outbreaks.
Therefore,
there
is
an
urgent
need
for
highly
sensitive
specific
approaches
identify
quantify
pathogens.
Clustered
Regularly
Interspaced
Short
Palindromic
Repeats
CRISPR/Cas
systems
Argonaute
(Ago)
belong
Microbial
Defense
Systems
(MDS).
The
guided,
programmable,
targeted
activation
nucleases
by
both
them
leading
way
new
generation
detection.
We
compare
these
two
in
terms
similarities
differences.
In
addition,
we
discuss
future
challenges
prospects
development
biosensors,
especially
electrochemical
biosensors.
This
review
expected
afford
researchers
entering
this
multidisciplinary
field
useful
guidance
provide
inspiration
more
innovative
biosensors
ACS Sensors,
Год журнала:
2021,
Номер
6(8), С. 2920 - 2927
Опубликована: Июль 19, 2021
Pathogenic
bacteria
infection
severely
threatens
human
health
and
causes
substantial
medical
financial
concern.
Rapid,
sensitive,
specific,
reliable
detection
of
pathogenic
is
crucial.
In
the
current
study,
a
CRISPR-Cas12a-powered
dual-mode
biosensor
was
developed
for
ultrasensitive
cross-validating
bacteria.
Simply,
amplicons
Salmonella
(used
as
model)-specific
invA
sequence
triggered
CRISPR-Cas12a-based
indiscriminate
degradation
single-stranded
DNAs
that
were
supposed
to
link
two
gold
nanoparticle
(AuNP)
probe
pairs,
inducing
an
aggregation-to-dispersion
change.
This
generated
observable
color
changes
became
even
more
apparent
after
centrifugation.
The
can
be
discerned
by
naked
eyes
recorded
using
portable
colorimeter.
Meanwhile,
photothermal
effect
CRISPR-Cas12-powered
AuNPs
explored
first
time
through
808
nm
near-infrared
irradiation
such
quantitative
measurement
carried
out
recording
temperatures
thermal
camera.
For
both
modes,
limit
1
CFU/mL
dynamic
range
from
100
108
obtained,
which
comparable
with
or
better
than
previously
reported
methods.
Our
proposed
demonstrated
satisfactory
selectivity
against
other
interfering
cells.
Furthermore,
this
proved
capable
in
food
samples.
Regarding
real
applications,
result
each
mode
used
cross-validation.
Only
case
having
doubly
confirmed
positive
negative
results
assured,
rendered
dependable
biosensing
conclusion.
technology
not
only
expands
reach
CRISPR-Cas
system
but
also
provides
general
method
sensing
desirable
sensitivity,
specificity,
capacity.
Analytical Chemistry,
Год журнала:
2023,
Номер
95(6), С. 3332 - 3339
Опубликована: Янв. 30, 2023
Herein,
a
chemiluminescence
(CL)
biosensor
based
on
CRISPR-Cas12a
and
cation
exchange
reaction
was
constructed
to
detect
the
biomarker
microRNA-21
(miRNA-21).
The
rolling
circle
amplification
(RCA)
introduced
convert
each
target
RNA
strand
into
long
single-stranded
DNA
with
repeated
sequences,
which
acted
as
triggers
initiate
transcleavage
activity
of
CRISPR-Cas12a.
activated
Cas12a
could
cleave
biotinylated
linker
CuS
nanoparticles
(NPs)
inhibit
binding
NPs
streptavidin
immobilized
surface
microplate,
strongly
reduced
generation
Cu2+
from
between
AgNO3,
thus
efficiently
suppressed
CL
Cu2+-luminol-H2O2
system,
giving
"signal
off"
biosensor.
With
multiple
amplification,
detection
limit
developed
sensor
for
miRNA-21
reached
16
aM.
In
addition,
this
is
not
only
suitable
professional
instrument
but
also
smartphone
used
tool
purpose
portable
low-cost
assay.
This
method
be
specifically
quite
low
level
in
human
serum
samples
various
cancer
cells,
indicating
its
potential
ultrasensitive
molecular
diagnostics.
Nature Communications,
Год журнала:
2024,
Номер
15(1)
Опубликована: Март 5, 2024
Abstract
Control
of
CRISPR/Cas12a
trans
-cleavage
is
crucial
for
biosensor
development.
Here,
we
show
that
small
circular
DNA
nanostructures
which
partially
match
guide
RNA
sequences
only
minimally
activate
Cas12a
ribonucleoproteins.
However,
linearizing
these
structures
restores
activation.
Building
on
this
finding,
an
Autocatalytic
Circular
Amplification
Reaction
(AutoCAR)
system
established
allows
a
single
nucleic
acid
target
to
multiple
ribonucleoproteins,
and
greatly
increases
the
achievable
reporter
cleavage
rates
per
target.
A
rate-equation-based
model
explains
observed
near-exponential
rate
trends.
Autocatalysis
also
sustained
with
modified
fluorophore-quencher
pairs
achieving
1
aM
level
(<1
copy/μL)
detection
(10
6
times
improvement),
without
additional
amplification,
within
15
min,
at
room
temperature.
The
range
tuneable,
spanning
3
11
orders
magnitude.
We
demonstrate
SNP
mutations
in
circulating
tumor
from
blood
plasma,
genomic
(
H.
Pylori
)
(SARS-CoV-2)
reverse
transcription
as
well
colorimetric
lateral
flow
tests
cancer
~100
sensitivity.
Biosensors,
Год журнала:
2023,
Номер
13(2), С. 202 - 202
Опубликована: Янв. 29, 2023
With
the
move
of
molecular
tests
from
diagnostic
labs
to
on-site
testing
becoming
more
common,
there
is
a
sudden
rise
in
demand
for
nucleic
acid-based
tools
that
are
selective,
sensitive,
flexible
terrain
changes,
and
cost-effective
assist
point-of-care
systems
large-scale
screening
be
used
remote
locations
cases
outbreaks
pandemics.
CRISPR-based
biosensors
comprise
promising
new
approach
acid
detection,
which
uses
Cas
effector
proteins
(Cas9,
Cas12,
Cas13)
as
extremely
specialized
identification
components
may
conjunction
with
variety
readout
approaches
(such
fluorescence,
colorimetry,
potentiometry,
lateral
flow
assay,
etc.)
onsite
analysis.
In
this
review,
we
cover
some
technical
aspects
integrating
CRISPR
system
traditional
biosensing
methods
amplification
technologies
such
polymerase
chain
reaction
(PCR),
loop-mediated
isothermal
(LAMP),
recombinase
(RPA)
continue
elaborate
on
prospects
developed
biosensor
detection
major
viral
bacterial
diseases.
Within
scope
article,
also
discuss
recent
COVID
pandemic
numerous
have
undergone
development
since
its
advent.
Finally,
challenges
future
testing.
