Sensors & Diagnostics,
Год журнала:
2024,
Номер
3(8), С. 1310 - 1318
Опубликована: Янв. 1, 2024
MicroRNAs
(miRNAs)
are
short
(about
18-24
nucleotides)
non-coding
RNAs
and
have
emerged
as
potential
biomarkers
for
various
diseases,
including
cancers.
Due
to
their
lengths,
the
specificity
often
becomes
an
issue
in
conventional
amplification-based
methods.
Next-generation
sequencing
techniques
could
be
alternative,
but
long
analysis
time
expensive
costs
make
them
less
suitable
routine
clinical
diagnosis.
Therefore,
it
is
essential
develop
a
rapid,
selective,
accurate
miRNA
detection
assay
using
simple,
affordable
system.
In
this
work,
we
report
CRISPR/Cas13a-based
biosensing
point-of-care
dark-field
(DF)
imaging.
We
utilized
magnetic-gold
nanoparticle
(MGNPs)
complexes
signal
probes,
which
consist
of
200
nm-sized
magnetic
beads
60
gold
nanoparticles
(AuNPs)
linked
by
DNA
hybridization.
Once
CRISPR/Cas13a
system
recognized
target
miRNAs
(miR-21-5p),
activated
Cas13a
cleaved
bridge
linker
containing
RNA
sequences,
releasing
nm-AuNPs
detected
quantified
portable
DF
imaging
The
combination
CRISPR/Cas13a,
MGNPs,
demonstrated
amplification-free
miR-21-5p
within
30
min
at
limit
500
attomoles
(25
pM)
with
single-base
specificity.
CRISPR/Cas13a-assisted
MGNP-DF
achieved
simple
equipment,
thus
providing
application
cancer
Analytical Chemistry,
Год журнала:
2022,
Номер
94(45), С. 15908 - 15914
Опубликована: Ноя. 3, 2022
The
current
CRISPR/Cas12a-based
diagnostic
techniques
focus
on
designing
the
crRNA
or
substrate
DNA
elements
to
indirectly
switch
trans-cleavage
activity
of
Cas12a
responsive
target
information.
Here,
we
propose
use
an
allosteric
probe
directly
regulate
and
present
a
method
for
sensing
different
types
analytes.
An
inhibitor
is
rationally
designed
couple
recognition
sequence
with
inhibitory
aptamer
CRISPR/Cas12a
system
enables
binding
specific
induce
change
conformation,
which
leads
loss
its
function
Cas12a.
As
result,
structure-switchable
can
degree
depending
dose
target.
Scalability
our
strategy
be
achieved
by
simply
replacing
loop
domain
sequences.
proposed
was
validated
detecting
adenosine
triphosphate
let-7a,
giving
detection
limits
490
nM
26
pM,
respectively,
showing
excellent
specificity.
We
believe
that
this
work
exploits
viable
approach
as
regulatory
element
biosensing
purposes,
enriching
arsenal
methods
molecular
diagnostics
spurring
further
development
application
aptamers
CRISPR/Cas
system.
Chemical Communications,
Год журнала:
2023,
Номер
59(50), С. 7763 - 7766
Опубликована: Янв. 1, 2023
In
this
work,
we
present
a
highly
sensitive,
specific,
and
versatile
method
to
quantify
miRNA
expression
by
coupling
CRISPR-Cas12a
with
cyclic
reverse
transcription
(CRT),
termed
as
CRISPR-CRT.
Each
target
was
first
converted
amplified
into
multiple
hairpin
RT
products
via
CRT.
Afterward,
the
could
serve
activators
initiate
collateral
cleavage
activity
of
CRISPR-Cas12a.
Due
above
two-stage
amplification,
assay
detect
at
sub-femtomolar
level
(LOD,
0.201
fM).
Since
sequence
is
double
checked:
in
CRT
then
CRISPR
system,
proposed
also
shows
an
excellent
specificity
detecting
miR-21.
Finally,
usage
assay,
sensitive
assessment
miR-21
levels
human
serum
samples
has
been
achieved
disease
detected.
Conclusively,
CRISPR-CRT
holds
great
application
prospective
field
clinical
molecular
diagnosis.
Chemical Communications,
Год журнала:
2024,
Номер
60(23), С. 3098 - 3117
Опубликована: Янв. 1, 2024
DNA
nanostructures
are
easy
to
design
and
construct,
have
good
biocompatibility,
show
great
potential
in
biosensing
drug
delivery.
Numerous
distinctive
versatile
been
developed
explored
for
biomedical
applications.
In
addition
that
completely
assembled
from
DNA,
composite
obtained
by
combining
with
other
organic
or
inorganic
materials
also
widely
used
related
research.
The
CRISPR/Cas
system
has
attracted
attention
as
a
powerful
gene
editing
technology
is
diagnosis.
Many
researchers
committed
exploring
new
possibilities
systems.
These
explorations
provide
support
the
development
of
detection
methods
cargo
delivery
pathways,
inspiration
improving
relevant
platforms,
further
expand
application
scope
This
paper
mainly
reviews
principles
applications
combined
based
on
types
nanostructures.
Finally,
status,
challenges
prospects
summarized.
It
expected
this
review
will
enable
better
understand
current
state
field
insights
into
systems
As
promising
noninvasive
biomarkers,
nucleic
acids
provide
great
potential
to
innovate
cancer
early
detection
methods
and
promote
subsequent
diagnosis
improve
the
survival
rates
of
patient.
Accurate,
straightforward
sensitive
such
acid-based
biomarkers
in
complex
biological
samples
holds
significant
clinical
importance.
However,
low
abundance
creates
huge
challenges
for
their
routine
detection.
next-generation
diagnostic
tool,
Clustered
Regularly
Interspaced
Short
Palindromic
Repeats
(CRISPR)/CRISPR-associated
protein
(Cas)
with
high
programmability,
sensitivity,
fidelity,
single-base
resolution,
precise
acid
positioning
capabilities
are
extremely
attractive
trace
(NABCBs),
permitting
rapid,
ultra-sensitive
specific
More
importantly,
by
combing
nanotechnology,
it
can
solve
long-lasting
problems
poor
accuracy
simplicity,
as
well
achieve
integrated
miniaturization
portable
point-of-care
testing
(POCT)
existing
literature
lacks
emphasis
on
this
topic.
Thus,
we
intend
propose
a
timely
one
readers.
This
review
will
bridge
gap
providing
insights
CRISPR/Cas-based
nano-biosensing
development
highlighting
current
state-of-art,
challenges,
prospects.
We
expect
that
better
understanding
valuable
NABCBs
detection,
thereby
facilitating
advancements
screening/detection/diagnostics
win
practical
applications
foreseeable
future.
