The rise and future of CRISPR-based approaches for high-throughput genomics
FEMS Microbiology Reviews,
Год журнала:
2024,
Номер
48(5)
Опубликована: Июль 31, 2024
Clustered
regularly
interspaced
short
palindromic
repeats
(CRISPR)
has
revolutionized
the
field
of
genome
editing.
To
circumvent
permanent
modifications
made
by
traditional
CRISPR
techniques
and
facilitate
study
both
essential
nonessential
genes,
interference
(CRISPRi)
was
developed.
This
gene-silencing
technique
employs
a
deactivated
Cas
effector
protein
guide
RNA
to
block
transcription
initiation
or
elongation.
Continuous
improvements
better
understanding
mechanism
CRISPRi
have
expanded
its
scope,
facilitating
genome-wide
high-throughput
screens
investigate
genetic
basis
phenotypes.
Additionally,
emerging
CRISPR-based
alternatives
further
possibilities
for
screening.
review
delves
into
CRISPRi,
compares
it
with
other
gene-perturbation
techniques,
highlights
superior
capacities
studying
complex
microbial
traits.
We
also
explore
evolution
emphasizing
enhancements
that
increased
capabilities,
including
multiplexing,
inducibility,
titratability,
predictable
knockdown
efficacy,
adaptability
nonmodel
microorganisms.
Beyond
we
discuss
activation,
RNA-targeting
systems,
single-nucleotide
resolution
perturbation
their
potential
in
Collectively,
this
gives
comprehensive
overview
general
workflow
screen,
an
extensive
discussion
strengths
weaknesses,
future
directions,
alternatives.
Язык: Английский
Accelerating enzyme discovery and engineering with high-throughput screening
Natural Product Reports,
Год журнала:
2024,
Номер
unknown
Опубликована: Янв. 1, 2024
Recent
progress
in
the
DBTL
cycle,
including
machine
learning,
facilitated
enzyme
mining
for
biocatalysis.
Automation
and
standardization
of
library
construction,
coupled
to
high-throughput
screening,
further
accelerates
discovery
process.
Язык: Английский
A Plasmid System That Utilises Phosphoribosylanthranilate Isomerase to Select Against Cells Expressing Truncated Proteins
Biomolecules,
Год журнала:
2025,
Номер
15(3), С. 412 - 412
Опубликована: Март 14, 2025
We
have
generated
a
vector
that
enables
the
removal
of
plasmids
coding
for
truncated
proteins.
This
expresses
protein
interest
in
yeast
Saccharomyces
cerevisiae
from
galactose-inducible
promoter.
The
gene
is
fused
in-frame
to
downstream
sequence
phosphoribosylanthranilate
isomerase
(PRAI),
which
catalyses
third
step
tryptophan
biosynthesis.
As
consequence,
only
full-length
renders
host
cell
prototrophic,
allowing
selection
against
cells
expressing
Our
proof-of-principle
study
demonstrates
PRAI
functional
when
C-terminally
protein,
robustly
rendering
prototrophic.
N-terminal
GST
tag
and
C-terminal
myc
allow
tag-mediated
purification,
co-precipitation
studies,
determination
relative
expression
levels,
as
well
validation
via
Western
blotting.
Язык: Английский
Industrial Microbial Technologies for Feed Protein Production from Non-Protein Nitrogen
Microorganisms,
Год журнала:
2025,
Номер
13(4), С. 742 - 742
Опубликована: Март 25, 2025
Due
to
the
increasing
global
demand
for
feed
protein,
microbial
protein
has
great
potential
of
being
able
sustainably.
However,
application
in
animal
cultivation
industry
is
still
limited
by
its
high
cost
and
availability
on
scale.
From
viewpoint
industrial
production,
it
vital
specify
crucial
processes
components
further
technical
exploration
process
optimization.
This
article
presents
state-of-the-art
technologies
non-protein
nitrogen
(NPN)
assimilation
production.
Nitrogen
sources
are
one
main
factors
media
used
large-scale
fermentation.
Therefore,
available
NPN
synthesis,
utilization
mechanisms,
fermentation
corresponding
strain
reviewed
this
paper.
Especially,
random
mutagenesis
adaptive
laboratory
evolution
(ALE)
approach
combined
with
(ultra-)
throughput
screening
provided
impetus
increase
yield.
Despite
underlying
technological
advances
production
extensive
research
development
efforts
required
before
commercial
feed.
Язык: Английский
Deaminase-Driven Random Mutation Enables Efficient DNA Mutagenesis for Protein Evolution
Chemical Science,
Год журнала:
2025,
Номер
unknown
Опубликована: Янв. 1, 2025
Protein
evolution
has
emerged
as
a
crucial
tool
for
generating
proteins
with
novel
characteristics.
A
key
step
in
protein
is
the
mutagenesis
of
protein-coding
DNA.
Error-prone
PCR
(epPCR)
is...
Язык: Английский
Adaptive laboratory evolution for improving acetogen gas fermentation
Current Opinion in Biotechnology,
Год журнала:
2025,
Номер
93, С. 103305 - 103305
Опубликована: Апрель 22, 2025
Gas
fermentation
using
acetogens
can
help
humankind
transition
from
petroleum-based
industries
to
more
sustainable
alternatives.
Acetogens
are
a
unique
set
of
organisms
that
efficiently
convert
carbon
oxide
waste
gases
into
chemicals,
such
as
ethanol
and
acetate.
While
already
used
in
commercially
operated
bioprocess
facilities,
the
field
is
still
affected
by
challenging
genetic
manipulation
workflows
developing
knowledge
acetogen
metabolism.
Adaptive
laboratory
evolution
(ALE)
uniquely
contribute
here,
through
guided
synthetically
created
niches,
which
delivers
strains
with
industrially
relevant
phenotypes
helps
resolve
genotype-phenotype
relationships.
Here,
we
review
expanding
use
ALE
for
acetogens,
showcasing
results
regarding
fundamental
understanding
improvement
-
faster
growth/substrate
utilisation,
elimination
media
components,
improving
stress
tolerance,
growth
robustness
bioreactor
cultures.
These
works
provide
opportunities
further
engineer
manipulate
traits
industrial
bioprocesses
improve
Язык: Английский
Engineering artificial microbial consortia for personalized gut microbiome modulation and disease treatment
Annals of the New York Academy of Sciences,
Год журнала:
2025,
Номер
unknown
Опубликована: Май 5, 2025
Abstract
The
human
gut
microbiome
is
a
complex
ecosystem
that
plays
vital
role
in
maintaining
health
and
contributing
to
the
pathogenesis
of
various
diseases.
This
review
proposes
transformative
approach
involves
engineering
artificial
microbial
consortia—precisely
designed
communities
microorganisms—for
personalized
modulation
targeted
therapeutic
interventions.
By
integrating
synthetic
biology,
systems
advanced
culturing
techniques,
tailored
consortia
can
be
developed
perform
specific
functions
within
gut,
including
production
molecules,
immune
responses,
competition
against
pathogenic
bacteria.
In
vitro
vivo
studies
indicate
these
engineered
effectively
restore
balance
enhance
host
resilience.
holds
immense
potential
revolutionize
healthcare
by
addressing
root
causes
diseases
such
as
metabolic
disorders,
inflammatory
conditions,
gastrointestinal
infections
through
precise
manipulation
microbiome.
Future
research
should
focus
on
rigorous
clinical
trials
evaluate
safety,
efficacy,
long‐term
impacts
diverse
populations,
paving
way
for
innovative
therapies
promote
overall
well‐being.
Язык: Английский
Beyond CEN.PK - parallel engineering of selected S. cerevisiae strains reveals that superior chassis strains require different engineering approaches for limonene production
Metabolic Engineering,
Год журнала:
2025,
Номер
unknown
Опубликована: Май 1, 2025
Язык: Английский
An in vivo target mutagenesis system for multiple hosts
Trends in biotechnology,
Год журнала:
2025,
Номер
unknown
Опубликована: Май 1, 2025
Язык: Английский
UV-Induced DNA Repair Mechanisms and Their Effects on Mutagenesis and Culturability in Escherichia coli
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2024,
Номер
unknown
Опубликована: Ноя. 14, 2024
Mutagenic
processes
drive
evolutionary
progress,
with
ultraviolet
(UV)
radiation
significantly
affecting
evolution.
Despite
extensive
research
on
SOS
response-mediated
mutagenesis,
UV-induced
repair
mechanisms
remain
complex,
and
their
effects
cell
survival
mutagenesis
are
not
fully
understood.
We
previously
observed
a
near-perfect
correlation
between
RecA-mediated
response
mutation
levels
in
Язык: Английский