bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2025,
Номер
unknown
Опубликована: Фев. 21, 2025
Abstract
Plasmids
remain
by
far
the
most
common
medium
for
delivering
engineered
DNA
to
microorganisms.
However,
reliance
on
natural
plasmid
replication
mechanisms
limits
their
tunability,
compatibility,
and
modularity.
Here
we
refactor
pMB1
origin
create
plasmids
with
customizable
copy
numbers
tuning
refactored
components.
We
then
compatible
origins
that
use
synthetic
RNA
regulators
implement
independent
control.
further
demonstrate
of
(SynORI)
can
be
modularly
respond
various
signals,
allowing
multiplexed
copy-based
reporting
environmental
signals.
Lastly,
a
library
6
orthogonal
SynORI
is
created
co-maintained
in
E.
coli
week.
This
work
establishes
feasibility
creating
serve
as
new
biotechnology
biology.
Plant Biotechnology Journal,
Год журнала:
2023,
Номер
21(7), С. 1440 - 1453
Опубликована: Апрель 9, 2023
Previous
work
has
demonstrated
that
plants
can
be
used
as
production
platforms
for
molecules
in
health,
medicine,
and
agriculture.
Production
been
exemplified
both
stable
transgenic
using
transient
expression
strategies.
In
particular,
species
of
Nicotiana
have
engineered
to
produce
a
range
useful
molecules,
including
insect
sex
pheromones,
which
are
valued
species-specific
control
agricultural
pests.
To
date,
most
studies
relied
on
strong
constitutive
all
pathway
genes.
However,
microbes
yields
improved
by
controlling
balancing
gene
expression.
Synthetic
regulatory
elements
provide
over
the
timing
levels
therefore
maximizing
from
heterologous
biosynthetic
pathways.
this
study,
we
demonstrate
use
engineering
synthetic
genetic
Lepidopteran
pheromones
benthamiana.
We
copper
low-cost
molecule
tightly
regulated
inducible
Further,
show
how
construct
architecture
influences
relative
and,
consequently,
product
multigene
constructs.
compare
number
orthogonal
maximal
constructs
is
mediated
dCas9-based
transcriptional
activators.
The
approaches
here
new
insights
into
reconstruction
metabolic
pathways
plants.
Annual Review of Chemical and Biomolecular Engineering,
Год журнала:
2024,
Номер
15(1), С. 267 - 292
Опубликована: Апрель 10, 2024
Augmenting
cells
with
novel,
genetically
encoded
functions
will
support
therapies
that
expand
beyond
natural
capacity
for
immune
surveillance
and
tissue
regeneration.
However,
engineering
at
scale
transgenic
cargoes
remains
a
challenge
in
realizing
the
potential
of
cell-based
therapies.
In
this
review,
we
introduce
range
applications
primary
stem
We
highlight
tools
advances
have
launched
mammalian
cell
from
bioproduction
to
precision
editing
therapeutically
relevant
cells.
Additionally,
examine
how
transgenesis
methods
genetic
cargo
designs
can
be
tailored
performance.
Altogether,
offer
vision
accelerating
translation
innovative
by
harnessing
diverse
types,
integrating
expanding
array
synthetic
biology
tools,
building
cellular
through
advanced
genome
writing
techniques.
Nucleic Acids Research,
Год журнала:
2024,
Номер
52(3), С. 1512 - 1521
Опубликована: Янв. 2, 2024
Abstract
The
field
of
synthetic
biology
and
biosystems
engineering
increasingly
acknowledges
the
need
for
a
holistic
design
approach
that
incorporates
circuit-host
interactions
into
process.
Engineered
circuits
are
not
isolated
entities
but
inherently
entwined
with
dynamic
host
environment.
One
such
interaction,
‘growth
feedback’,
results
when
modifications
in
growth
patterns
influence
operation
gene
circuits.
growth-mediated
effects
can
range
from
growth-dependent
elevation
protein/mRNA
dilution
rate
to
changes
resource
reallocation
within
cell,
which
lead
complete
functional
collapse
complex
To
achieve
robust
circuit
performance,
biologists
employ
variety
control
mechanisms
stabilize
insulate
behavior
against
changes.
Here
we
propose
simple
strategy
by
incorporating
one
repressive
edge
growth-sensitive
bistable
circuit.
Through
both
simulation
vitro
experimentation,
demonstrate
how
this
additional
node
stabilizes
protein
levels
increases
robustness
response
feedback.
We
incorporation
links
as
desensitizing
fluctuations.
ACS Synthetic Biology,
Год журнала:
2022,
Номер
12(1), С. 27 - 34
Опубликована: Дек. 14, 2022
Vinblastine
is
a
chemotherapy
agent
produced
by
the
plant
Catharanthus
roseus
in
small
quantities.
Currently,
vinblastine
sourced
isolation
or
semisynthesis.
Nicotiana
benthamiana
heterologous
host
that
can
be
used
for
reconstitution
of
biosynthetic
pathways
as
an
alternative
natural
product
sourcing
strategy.
Recently,
biosynthesis
late-stage
precursors
precondylocarpine
acetate,
catharanthine,
and
tabersonine
have
been
fully
elucidated.
However,
large
number
enzymes
involved
pathway
unstable
nature
intermediates
make
precursor
challenging.
We
N.
chassis
state-of-art
modular
vector
assembly
to
optimize
six
steps
leading
production
acetate
from
central
intermediate
strictosidine
(∼2.7
mg
per
1
g
frozen
tissue).
After
selecting
optimal
regulatory
element
combination,
we
constructed
four
transcriptional
unit
assemblies
tested
their
efficiency.
Finally,
successfully
reconstituted
catharanthine
tabersonine.
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2024,
Номер
unknown
Опубликована: Окт. 17, 2024
Abstract
Targeting
DNA
payloads
into
human
(h)iPSCs
involves
multiple
time-consuming,
inefficient
steps
that
must
be
repeated
for
each
construct.
Here,
we
present
STRAIGHT-IN
Dual,
which
enables
simultaneous,
allele-specific,
single-copy
integration
of
two
with
100%
efficiency
within
one
week.
Notably,
Dual
leverages
the
platform
to
allow
near-scarless
cargo
integration,
facilitating
recycling
components
subsequent
cellular
modifications.
Using
investigated
how
promoter
choice
and
gene
syntax
influences
transgene
silencing,
demonstrate
impact
these
design
features
on
forward
programming
hiPSCs
neurons.
Furthermore,
designed
a
grazoprevir-inducible
synZiFTR
system
complement
widely-used
tetracycline-inducible
system,
providing
independent,
tunable,
temporally
controlled
expression
both
transcription
factors
functional
reporters.
The
unprecedented
speed
generates
homogenous
genetically
engineered
hiPSC
populations
represents
major
advancement
synthetic
biology
in
stem
cell
applications
opens
opportunities
precision
engineering.
Nature Communications,
Год журнала:
2024,
Номер
15(1)
Опубликована: Окт. 30, 2024
Abstract
During
development,
most
cells
experience
a
progressive
restriction
of
fate
that
ultimately
results
in
fully
differentiated
mature
state.
Understanding
more
about
the
gene
expression
patterns
underlie
developmental
programs
can
inform
engineering
efforts
for
new
or
optimized
forms.
Here,
we
present
four-state
integrase-based
recorder
history
and
demonstrate
its
use
tracking
events
Arabidopsis
thaliana
two
contexts:
lateral
root
initiation
stomatal
differentiation.
The
uses
serine
integrases
to
mediate
sequential
DNA
recombination
events,
resulting
step-wise,
history-dependent
switching
between
fluorescent
reporters.
By
using
promoters
express
at
different
times
along
each
differentiation
pathways
drive
integrase
expression,
tie
status
an
ordered
progression
trajectory.
In
one
snapshot
tissue,
our
is
able
reveal
past
with
single
cell
resolution.
this
way,
are
capture
heterogeneity
confirming
existence
alternate
paths
