ACS Sensors,
Journal Year:
2024,
Volume and Issue:
9(7), P. 3616 - 3624
Published: July 9, 2024
Clustered
Regularly
Interspaced
Short
Palindromic
Repeats-CRISPR-Associated
Protein
(CRISPR-Cas)
systems
have
evolved
several
mechanisms
to
specifically
target
foreign
DNA.
These
properties
made
them
attractive
as
biosensors.
The
primary
drawback
associated
with
contemporary
CRISPR-Cas
biosensors
is
their
weak
signaling
capacity,
which
typically
compensated
for
by
coupling
the
nucleic
acid
amplification.
An
alternative
strategy
improve
capacity
engineer
reporter,
i.e.,
design
new
signal-generating
substrates
Cas
proteins.
Unfortunately,
due
reliance
on
custom
synthesis,
most
of
these
engineered
reporter
are
inaccessible
many
researchers.
Herein,
we
investigate
a
substrate
based
fluorescein
(FAM)–tetramethylrhodamine
(TAMRA)
Förster
resonant
energy-transfer
(FRET)
pair
that
functions
seamless
"drop-in"
replacement
existing
reporters,
without
need
change
any
other
aspect
CRISPR-Cas12a-based
assay.
readily
available
and
employs
FRET
produce
two
signals
upon
cleavage
Cas12a.
use
both
in
ratiometric
manner
provides
improved
assay
performance
decreased
time-to-result
CRISPR-Cas12a
assays
when
compared
traditional
FAM–Black
Hole
Quencher
(BHQ)
quench-based
reporter.
We
comprehensively
characterize
this
better
understand
reasons
benchmark
it
against
current
standard
Finally,
showcase
real-world
utility
employ
Recombinase
Polymerase
Amplification
(RPA)–CRISPR-Cas12a
DNA
Endonuclease-Targeted
CRISPR
Trans
Reporter
(DETECTR)
detect
Human
papillomavirus
patient-derived
samples.
ACS Synthetic Biology,
Journal Year:
2024,
Volume and Issue:
13(6), P. 1633 - 1646
Published: June 11, 2024
A
growing
number
of
applications
require
simultaneous
detection
multiplexed
nucleic
acid
targets
in
a
single
reaction,
which
enables
higher
information
density
combination
with
reduced
assay
time
and
cost.
Clustered
regularly
interspaced
short
palindromic
repeats
(CRISPR)
the
CRISPR-Cas
system
have
broad
for
acids
due
to
their
strong
specificity,
high
sensitivity,
excellent
programmability.
However,
realizing
is
still
challenging
nonspecific
collateral
cleavage
activity,
limited
signal
reporting
strategies,
possible
cross-reactions.
In
this
review,
we
summarize
principles,
features
based
on
further
discuss
challenges
perspective.
Analytical Methods,
Journal Year:
2024,
Volume and Issue:
unknown
Published: Jan. 1, 2024
Reviews
the
development
of
visual
detection
methods
in
recent
years
based
on
different
Cas
and
analyzes
their
advantages
disadvantages
as
well
challenges
future
research.
Bioconjugate Chemistry,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Jan. 21, 2025
Erythropoietin
(EPO)-induced
cellular
signaling
through
the
EPO
receptor
(EPOR)
is
a
fundamental
pathway
for
modulation
of
behavior
and
activity.
In
our
previous
work,
we
showed
in
primary
human
astrocytes
that
multivalent
display
on
surface
semiconductor
quantum
dots
(QDs)
mediates
augmented
JAK/STAT
signaling,
concomitant
1.8-fold
increase
expression
aquaporin-4
(AQPN-4)
channel
proteins,
2-fold
AQPN-4-mediated
water
transport
Our
hypothesis
this
enhanced
involves
simultaneous
ligation
clustering
EPOR
by
QD-EPO
conjugates.
Here,
utilized
embryonic
kidney
(HEK
293T/17)
cell
line
transfected
with
fused
to
green
fluorescent
protein
(eGFP)
visualize
clustering.
We
demonstrate
QDs
displaying
five
copies
(bearing
C-terminal
6-histidine
tract)
nanoparticle
induce
compared
monomeric
at
same
concentration.
findings
confirm
critical
role
played
mediating
EPOR.
More
generally,
these
results
illustrate
capability
nanoparticle-growth
factor
bioconjugates
control
activity
cognate
receptors
important
selective
delivery
signaling.
Lab on a Chip,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Jan. 1, 2025
Smartphones
have
the
potential
to
transform
chemical
and
biological
analysis
by
functioning
as
portable,
versatile,
democratized
platforms
for
molecular
diagnostics.
Recent
advances
in
optical
detection
devices
are
reviewed.
Biosensors and Bioelectronics,
Journal Year:
2025,
Volume and Issue:
279, P. 117402 - 117402
Published: March 31, 2025
CRISPR-based
diagnostics
have
gained
increasing
attention
as
biosensing
tools
able
to
address
limitations
in
contemporary
molecular
diagnostic
tests.
To
maximize
the
performance
of
assays,
much
effort
has
focused
on
optimizing
chemistry
and
biology
reaction.
However,
less
been
paid
improving
techniques
used
analyze
data.
date,
decisions
typically
involve
various
forms
slope-based
classification.
Such
methods
are
superior
traditional
based
assessing
absolute
signals,
but
still
limitations.
Herein,
we
establish
benchmarks
(total
accuracy,
sensitivity,
specificity)
using
common
methods.
We
compare
these
benchmark
with
three
different
quadratic
empirical
distribution
function
statistical
tests,
finding
significant
improvements
speed
accuracy
when
applied
a
clinical
data
set.
Two
techniques,
Kolmogorov-Smirnov
Anderson-Darling
report
lowest
time-to-result
highest
total
test
accuracy.
Furthermore,
developed
long
short-term
memory
recurrent
neural
network
classify
CRISPR-biosensing
data,
achieving
100
%
specificity
our
model
Finally,
provide
guidelines
choosing
classification
method
parameters
that
best
suit
assay's
needs.
