ACS Sensors,
Journal Year:
2023,
Volume and Issue:
8(11), P. 3988 - 4007
Published: Oct. 23, 2023
Point-of-care
(POC)
detection
is
getting
more
and
attention
in
many
fields
due
to
its
accuracy
on-site
test
property.
The
CRISPR/Cas12a
system
endowed
with
excellent
sensitivity,
target
identification
specificity,
signal
amplification
ability
biosensing
because
of
unique
trans-cleavage
ability.
As
a
result,
lot
research
has
been
made
develop
CRISPR/Cas12a-based
biosensors.
In
this
review,
we
focused
on
readout
strategies
summarized
recent
sensitivity-improving
fluorescence,
colorimetric,
electrochemical
signaling.
Then
introduced
novel
portability-improving
based
lateral
flow
assays
(LFAs),
microfluidic
chips,
simplified
instruments,
one-pot
design.
the
end,
also
provide
our
outlook
for
future
development
Chemical Science,
Journal Year:
2022,
Volume and Issue:
13(15), P. 4364 - 4371
Published: Jan. 1, 2022
Besides
gene-editing,
the
CRISPR/Cas12a
system
has
also
been
widely
used
in
vitro
biosensing,
but
its
applications
live-cell
biosensing
are
rare.
One
reason
is
lacking
appropriate
carriers
to
synchronously
deliver
all
components
of
into
living
cells.
Herein,
we
demonstrate
that
MnO2
nanosheets
an
excellent
carrier
due
two
important
roles
played
by
them.
Through
a
simple
mixing
operation,
can
be
loaded
on
and
thus
delivered
Intracellular
glutathione
(GSH)-induced
decomposition
not
only
results
rapid
release
cells
provides
Mn2+
as
accelerator
promote
CRISPR/Cas12a-based
intracellular
targets.
Due
merits
highly
efficient
delivery,
release,
accelerated
signal
output
reaction,
work
better
than
commercial
liposome
analysis
survivin
messenger
RNA
(mRNA),
producing
much
brighter
fluorescence
images
shorter
time.
The
use
might
provide
good
for
different
CRISPR/Cas
systems
achieve
sensitive
targets,
paving
promising
way
ACS Nano,
Journal Year:
2022,
Volume and Issue:
16(12), P. 20693 - 20704
Published: Nov. 15, 2022
Strategies
utilizing
the
CRISPR/Cas
nucleases
Cas13
and
Cas12
have
shown
great
promise
in
development
of
highly
sensitive
rapid
diagnostic
assays
for
detection
pathogenic
nucleic
acids.
The
most
common
approaches
fluorophore-quencher
molecular
beacons
require
strand
amplification
strategies
or
optical
setups
to
overcome
limitations
readout.
Here,
we
demonstrate
a
flexible
strategy
assembling
luminescent
colorimetric
quantum
dot-nucleic
acid
hairpin
(QD-HP)
use
diagnostics.
This
utilizes
chimeric
peptide-peptide
(peptide-PNA)
conjugate
fluorescently
labeled
DNA
RNA
hairpins
ZnS-coated
QDs.
QDs
are
particularly
promising
alternatives
due
their
greater
brightness,
strong
UV
absorbance
with
large
emission
offset,
exceptional
photostability,
potential
multiplexing
sharp
peaks.
Using
Förster
resonance
energy
transfer
(FRET),
developed
ratiometric
reporters
capable
pM
target
(without
nucleotide
amplification)
both
RNA,
further
demonstrated
capabilities
camera-phone
detection.
flexibility
this
system
is
imparted
by
dual
functionality
QD
as
FRET
donor
central
nanoscaffold
arranging
acids
fluorescent
acceptors
on
its
surface.
method
also
provides
generalized
approach
that
could
be
applied
other
nuclease
systems.
ACS Sensors,
Journal Year:
2023,
Volume and Issue:
8(7), P. 2852 - 2858
Published: July 4, 2023
Rapid
and
accurate
detection
of
biomarkers
was
very
important
for
early
screening
treatment
diseases.
Herein,
a
sensitive
amplification-free
electrochemiluminescence
(ECL)
biosensor
based
on
CRISPR/Cas12a
DNA
tetrahedron
nanostructures
(TDNs)
constructed.
Briefly,
3D
TDN
self-assembled
the
Au
nanoparticle-deposited
glassy
carbon
electrode
surface
to
construct
biosensing
interface.
The
presence
target
would
activate
trans-cleavage
activity
Cas12a-crRNA
duplex
cleave
single-stranded
signal
probe
vertex
TDN,
causing
Ru(bpy)32+
fall
from
weakened
ECL
signal.
Thus,
system
transduced
change
concentration
into
an
enabling
HPV-16.
specific
recognition
HPV-16
made
have
good
selectivity,
while
TDN-modified
sensing
interface
could
reduce
cleaving
steric
resistance
improve
performance
CRISPR/Cas12a.
In
addition,
pretreated
complete
sample
within
100
min
with
limit
8.86
fM,
indicating
that
developed
possesses
potential
application
prospect
fast
nucleic
acid
detection.
Small,
Journal Year:
2023,
Volume and Issue:
19(49)
Published: Aug. 23, 2023
Abstract
Digital
nucleic
acid
detection
based
on
microfluidics
technology
can
quantify
the
initial
amount
of
in
sample
with
low
equipment
requirements
and
simple
operations,
which
be
widely
used
clinical
vitro
diagnosis.
Recently,
isothermal
amplification
technologies
such
as
recombinase
polymerase
(RPA),
loop‐mediated
(LAMP),
clustered
regularly
interspaced
short
palindromic
repeats‐CRISPR
associated
proteins
(CRISPR‐Cas)
assisted
have
become
a
hot
spot
attention
state‐of‐the‐art
digital
chips
provided
powerful
tool
for
these
technologies.
Herein,
including
RPA,
LAMP,
CRISPR‐Cas
methods,
recently,
been
reviewed.
Moreover,
challenges
possible
strategies
to
address
them
are
discussed.
Finally,
future
directions
technology,
microfluidic
chip
device
manufacturing,
multiplex
detection,
one‐pot
outlined.
TrAC Trends in Analytical Chemistry,
Journal Year:
2023,
Volume and Issue:
162, P. 117028 - 117028
Published: March 18, 2023
In
addition
to
its
remarkable
genome
editing
capability,
the
CRISPR-Cas
system
has
proven
be
very
effective
in
many
fields
of
application,
including
biosensing
pathogenic
infections,
mutagenic
defects,
or
early
cancer
diagnosis.
Thanks
their
advantages
terms
simplicity,
efficiency,
and
reduced
time,
several
systems
have
been
described
for
design
sensitive
selective
analytical
tools,
paving
way
development
further
commercialization
next-generation
diagnostics.
However,
CRISPR-Cas-based
biosensors
still
need
research
efforts
improve
some
drawbacks,
such
as
target
amplification,
low
reproducibility,
lack
knowledge
exploited
element
robustness.
This
review
aims
describe
latest
trends
technologies
better
highlight
insights
point
out
limitations
that
overcome
future
market
entry
medical
