Abstract
Currently,
with
the
growing
global
demand
for
infectious
disease
diagnostics,
environmental
testing
and
food
safety,
CRISPR‐Cas
(Clustered
Regularly
Interspaced
Short
Palindromic
Repeats‐CRISPR
associated
proteins)
system
has
unlimited
potential
as
one
of
emerging
technologies
in
field
biotechnology.
Recent
studies
have
shown
that
become
a
powerful
weapon
gene
editing,
treatment,
molecular
diagnosis
imaging.
As
important
biomarkers,
various
nucleic
acid
non‐nucleic
targets
need
to
be
detected
from
samples
before
situation
deteriorates.
In
addition,
there
are
still
some
limitations
challenges
current
biosensor
platforms
based
on
system.
Therefore,
summarising
strategies,
strengths
weaknesses,
development
prospects
target
identification
is
beneficial
fully
explore
directions
CRISPR‐Cas‐based
systems.
This
review
briefly
introduces
summarises
classification,
mechanisms
applications
(diagnostics
when
combined
signal
detection
means).
limitations,
future
optimisation
this
discussed.
Critical Reviews in Analytical Chemistry,
Год журнала:
2024,
Номер
unknown, С. 1 - 17
Опубликована: Март 15, 2024
MicroRNA
(miRNA)
has
emerged
as
a
promising
biomarker
for
disease
diagnosis
and
potential
therapeutic
targets
drug
development.
The
detection
of
miRNA
can
serve
noninvasive
tool
in
diseases
predicting
prognosis.
CRISPR/Cas12a
system
great
nucleic
acid
due
to
its
high
sensitivity
specificity,
which
been
developed
be
versatile
acid-based
various
fields.
However,
conversion
from
RNA
DNA
with
or
without
amplification
operation
is
necessary
based
on
system,
because
dsDNA
containing
PAM
sequence
ssDNA
traditionally
considered
the
activator
Cas12a.
Until
recently,
direct
by
reported.
In
this
review,
we
provide
an
overview
evolution
biosensors
indirect
direct,
would
beneficial
development
CRISPR/Cas12a-based
sensors
better
performance
miRNA.
Biosensors,
Год журнала:
2023,
Номер
13(7), С. 747 - 747
Опубликована: Июль 20, 2023
MicroRNAs
(miRNAs)
are
a
class
of
small
noncoding
RNAs
that
approximately
22
nt
in
length
and
regulate
gene
expression
post-transcriptionally.
miRNAs
play
vital
role
both
physiological
pathological
processes
regarded
as
promising
biomarkers
for
cancer,
cardiovascular
diseases,
neurodegenerative
so
on.
Accurate
detection
miRNA
level
clinical
samples
is
important
miRNA-guided
diagnostics.
However,
the
common
approaches
like
RNA
sequencing,
qRT-PCR,
microarray
performed
professional
laboratory
with
complex
intermediate
steps
time-consuming
costly,
challenging
Hence,
sensitive,
highly
specific,
rapid,
easy-to-use
crucial
diagnosis
based
on
miRNAs.
With
advantages
being
efficient,
cost-saving,
easy
to
operate,
point-of-care
testing
(POCT)
has
been
widely
used
For
first
time,
we
mainly
focus
summarizing
research
progress
POCT
portable
instruments
visual
readout
methods.
As
available
pocket-size
roles
POCT,
provide
an
all-sided
discussion
principles
these
methods
their
main
limitations
challenges,
order
guide
development
more
accurate,
sensitive
detection.
Analytical Methods,
Год журнала:
2025,
Номер
unknown
Опубликована: Янв. 1, 2025
DNA
methyltransferase
(DNA
MTase)
is
a
valuable
target
of
genetic
diseases,
and
detection
related
MTase
activity
very
important
for
drug
screening,
clinical
diagnosis
disease
treatment.
Herein,
universal
two-step
strategy
based
on
CRISPR/Cas12a-assisted
hyperbranched
rolling
circle
amplification
(CA-HRCA)
constructed,
which
successfully
achieves
the
Dam
M.SssI
MTase.
In
presence
restriction
enzymes,
HRCA
primer
locked
in
dumbbell
probe
will
be
released
further
initiates
HRCA.
first
step,
methylation,
enzyme
digestion
are
performed
simultaneously,
effectively
simplifying
reaction
process
shortening
time.
second
abundant
products
(dsDNA)
act
as
activators
to
induce
CRISPR/Cas12a
split
fluorescent
probes.
Compared
with
ssDNA
activators,
dsDNA
can
cause
higher
collateral
cleavage
CRISPR/Cas12a.
As
expected,
this
presents
excellent
sensing
performance
time
155
min.
The
LODs
calculated
7.6
×
10-4
U
mL-1
1.8
mL-1,
respectively.
And
proposed
assay
possesses
extraordinary
specificity
reproducibility.
Moreover,
practical
application
ability
development
potential
proved
by
serum
spiked
test
inhibitor
evaluation
tests.
