Biomolecules,
Год журнала:
2023,
Номер
13(7), С. 1022 - 1022
Опубликована: Июнь 21, 2023
Programmable
endonucleases,
such
as
Cas
(Clustered
Regularly-Interspaced
Short
Repeats-associated
proteins)
and
prokaryotic
Argonaute
(pAgo),
depend
on
base
pairing
of
the
target
DNA
with
guide
RNA
or
to
cleave
strands.
Therefore,
they
are
capable
recognizing
cleaving
sequences
at
virtually
any
arbitrary
site.
The
present
review
focuses
commonly
used
in
vivo
vitro
recombination-based
gene
cloning
methods
application
programmable
endonucleases
these
sequence-
ligation-independent
assembly
methods.
advantages
shortcomings
utilized
tools
for
also
discussed
this
review.
Current Opinion in Microbiology,
Год журнала:
2023,
Номер
74, С. 102313 - 102313
Опубликована: Апрель 5, 2023
Both
eukaryotes
and
prokaryotes
(archaea
bacteria)
encode
an
arsenal
of
immune
systems
that
protect
the
host
against
mobile
genetic
elements
(MGEs)
including
viruses,
plasmids,
transposons.
Whereas
Argonaute
proteins
(Agos)
are
best
known
for
post-transcriptional
gene
silencing
in
eukaryotes,
all
domains
life,
members
from
highly
diverse
protein
family
act
as
programmable
systems.
To
this
end,
Agos
programmed
with
small
single-stranded
RNA
or
DNA
guides
to
detect
silence
complementary
MGEs.
Across
within
different
function
distinct
pathways
MGE
detection
can
trigger
various
mechanisms
provide
immunity.
In
review,
we
delineate
underlying
both
eukaryotic
Argonautes
(eAgos)
prokaryotic
(pAgos).
Analytical Chemistry,
Год журнала:
2024,
Номер
96(2), С. 620 - 623
Опубликована: Янв. 3, 2024
Prokaryotic
Argonautes
(pAgos)
have
been
recently
used
in
many
nucleic
acid
biosensing
applications
but
rarely
for
regulating
the
isothermal
amplification
system.
Herein,
we
reported
Thermus
thermophilus
Argonaute
(TtAgo)-mediated
background-suppressed
exponential
(EXPAR)
as
first
example
to
explore
binding
activity
of
pAgos
toward
regulation
template.
It
was
demonstrated
that
thermophilic
efficiently
eliminated
nonspecific
hybridization
between
templates
by
their
affinity
with
template,
resulting
greatly
enhancing
specificity
EXPAR.
TtAgo-mediated,
EXPAR
employed
detect
miRNA
a
detection
limit
10–15
M,
which
1000
times
and
100
more
sensitive
than
traditional
RT-PCR
EXPAR,
respectively.
This
method
further
showed
good
performance
discriminating
cancer
patients
from
healthy
individuals,
indicating
its
potential
practical
clinical
applications.
Short
prokaryotic
argonaute
(pAgo)
and
toll/interleukin-1
receptor/resistance
protein
(TIR)-analog
of
PAZ
(APAZ)
form
a
heterodimeric
SPARTA
complex
that
provides
immunity
to
its
host
through
an
abortive
infection
mechanism.
Monomeric
senses
foreign
RNA/DNA
duplexes
assemble
active
tetramer
resulting
in
cell
death
by
nicotinamide
adenine
dinucleotide
(oxidized
form)
(NAD)
depletion
via
unknown
We
report
nine
structures
different
functional
states
at
resolution
range
4.2
2.9
angstroms,
revealing
activation
Inactive
monomers
bind
symmetric
dimers
mediated
the
association
Ago
subunits.
The
initiation
assembly
induces
flexibility
TIR
domains
enabling
symmetry-breaking
rotational
movement
domain
dimer
units
which
facilitates
oligomerization,
formation
substrate
binding
pocket
complex's
NADase
activity.
Our
findings
provide
detailed
structural
mechanistic
insights
into
activating
short
defense
system.
Frontiers in Microbiology,
Год журнала:
2023,
Номер
14
Опубликована: Сен. 8, 2023
It
is
vital
to
diagnose
pathogens
quickly
and
effectively
in
the
research
treatment
of
disease.
Argonaute
(Ago)
proteins
are
recently
discovered
nucleases
with
nucleic
acid
shearing
activity
that
exhibit
specific
recognition
properties
beyond
CRISPR-Cas
nucleases,
which
highly
researched
but
restricted
PAM
sequence
recognition.
Therefore,
on
Ago
protein-mediated
detection
technology
has
attracted
significant
attention
from
researchers
recent
years.
Using
developing
platforms
can
enable
efficient,
convenient,
rapid
pathogen
diagnosis,
great
importance
for
human
life
health
technological
development.
In
this
article,
we
introduce
structure
function
discuss
latest
advances
their
use
detection.
Biosensors,
Год журнала:
2023,
Номер
13(9), С. 885 - 885
Опубликована: Сен. 15, 2023
Isothermal
nucleic
acid
amplification
tests
have
recently
gained
popularity
over
polymerase
chain
reaction
(PCR),
as
they
only
require
a
constant
temperature
and
significantly
simplify
amplification.
Recently,
numerous
attempts
been
made
to
incorporate
paper
microfluidics
into
these
isothermal
tests.
Paper
(including
lateral
flow
strips)
used
extract
acids,
amplify
the
target
gene,
detect
amplified
products,
all
toward
automating
process.
We
investigated
literature
from
2020
present,
i.e.,
since
onset
of
COVID-19
pandemic,
during
which
significant
surge
in
has
observed.
microfluidic
detection
extensively
for
recombinase
(RPA)
its
related
methods,
along
with
loop-mediated
(LAMP)
rolling
circle
(RCA).
Detection
was
conducted
primarily
colorimetric
fluorometric
although
few
publications
demonstrated
distance-
surface-enhanced
Raman
spectroscopic
(SERS)-based
detection.
A
good
number
could
be
found
that
both
on
platforms.
small
showed
extraction
or
three
procedures
(i.e.,
fully
integrated
systems)
platforms,
necessitating
need
future
work.
Biosensors,
Год журнала:
2023,
Номер
13(10), С. 941 - 941
Опубликована: Окт. 20, 2023
The
advancement
in
CRISPR-Cas
biosensors
has
transmuted
the
detection
of
plant
viruses
owing
to
their
rapid
and
higher
sensitivity.
However,
false
positives
restricted
multiplexing
capabilities
are
still
challenges
faced
by
this
technology,
demanding
exploration
novel
methodologies.
In
study,
a
system
was
developed
integrating
reverse
transcriptome
(RT)
techniques
with
recombinase
polymerase
isothermal
amplification
(RPA)
Frontiers in Bioscience-Landmark,
Год журнала:
2025,
Номер
30(2)
Опубликована: Фев. 14, 2025
Phages
have
exerted
severe
evolutionary
pressure
on
prokaryotes
over
billions
of
years,
resulting
in
major
rearrangements.
Without
every
enzyme
involved
the
phage–bacterium
interaction
being
examined;
bacteriophages
cannot
be
used
practical
applications.
Numerous
studies
conducted
past
few
years
uncovered
a
huge
variety
bacterial
antiphage
defense
systems;
nevertheless,
mechanisms
most
these
systems
are
not
fully
understood.
Understanding
interactions
between
bacteriophage
and
proteins
is
important
for
efficient
host
cell
infection.
Phage
bacteriophage–host
often
arise
immediately
after
Here,
we
review
main
groups
phage
enzymes
first
stage
viral
infection
responsible
degradation
membrane.
These
include
polysaccharide
depolymerases
(endosialidases,
endorhamnosidases,
alginate
lyases,
hyaluronate
lyases),
peptidoglycan
hydrolases
(ectolysins
endolysins).
Host
target
inhibited,
activated,
or
functionally
redirected
by
protein.
determine
bacteria.
Proteins
interest
holins,
endolysins,
spanins,
which
release
progeny
during
lytic
cycle.
This
describes
analyzes
therapeutic
potential
bacteriophage-derived
proteins.
