Analytical Chemistry,
Journal Year:
2024,
Volume and Issue:
96(42), P. 16735 - 16742
Published: Oct. 14, 2024
A
key
challenge
for
achieving
high-efficient
DNA
strand
displacement
reaction
(SDR)
with
existing
technologies
is
the
inferior
kinetic
performance
due
to
alternately
cumbersome
conjunction
and
dissociation
of
dsDNA.
In
this
work,
a
novel
multidriven
SDR
collaborated
by
toehold
initiator,
towing,
click
chemistry
engineered.
The
invasion
(O)
endows
hybridization
basal
(M)
in
dsDNA
releasing
(P),
which
can
be
significantly
boosted
towing
helper
impetus
from
reaction.
Accordingly,
rate
extent
P
largely
improved
showed
desiring
close
6-fold
compared
traditional
method,
providing
newly
strategy
potential
application
biosensing,
clinical
diagnostics,
nanotechnology.
view
this,
practical
biosensing
platform
combining
(MSDR)
waste-free
multi-cycle
amplification
constructed
rapid
ultrasensitive
electrochemical
detection
cancer-related
miRNA-21.
substantial
output
as
an
target-triggered
multicycle
high-efficiently
release
signal
probe
(Fc)-labeled
(P)
on
electrode
using
proposed
MSDR,
obtaining
low
limit
below
106.8
aM.
Analytical Chemistry,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Feb. 6, 2025
Despite
the
widespread
application
of
CRISPR-Cas12a
system
in
vitro
diagnostics
due
to
its
high
programmability
and
distinctive
trans-cleavage
activity,
susceptibility
crRNA
component
degradation
sensitivity
storage
working
conditions
poses
a
significant
challenge
improving
practical
efficacy
these
diagnostic
systems.
Here,
we
show
that
engineered
with
covalently
closed
circular
structure
(C-crRNA)
can
replace
traditional
linear
form
functional
complexes
Cas12a
protein,
significantly
enhancing
anti-interference
ability
while
maintaining
specificity.
Based
on
this
finding,
crRNA-mediated
CRISPR
molecular
(CRCD)
toolkit
is
developed
successfully
integrated
standard
nucleic
acid
amplification
technique
detect
synthesized
Human
Papillomavirus
type
16
(HPV-16)
plasmids
down
10
aM
levels.
Furthermore,
CRCD
applied
for
ultrasensitive
detection
40
HPV-16
influenza
A
viruses
clinical
samples,
results
consistent
those
from
PANTHER
quantitative
real-time
polymerase
chain
reaction
(qRT-PCR).
In
conclusion,
strategy
introduces
novel
paradigm
engineering
program
Cas12a,
which
has
potential
revolutionize
use
CRISPR-based
diagnostics.
