Navigating gene editing in porcine embryos: Methods, challenges, and future perspectives
Genomics,
Год журнала:
2025,
Номер
117(2), С. 111014 - 111014
Опубликована: Фев. 13, 2025
Gene
editing
technologies,
particularly
CRISPR/Cas9,
have
emerged
as
transformative
tools
in
genetic
modification,
significantly
advancing
the
use
of
porcine
embryos
biomedical
and
agricultural
research.
This
review
comprehensively
examines
various
methodologies
for
gene
delivery
methods,
such
somatic
cell
nuclear
transfer
(SCNT),
microinjection,
electroporation,
lipofection.
review,
focuses
on
advantages
or
limitations
using
different
biological
sources
(in
vivo-
vs.
vitro
oocytes/embryos).
Male
germ
manipulation
sperm-mediated
(SMGT)
testis-mediated
(TMGT)
represent
innovative
approaches
producing
genetically
modified
animals.
Although
these
technologies
revolutionized
engineering
field,
all
strategies
face
challenges,
including
high
rates
off-target
events
mosaicism.
emphasizes
need
to
refine
with
a
focus
reducing
mosaicism
improving
accuracy.
Further
advancements
are
essential
unlocking
full
potential
both
applications
innovations.
Язык: Английский
Optimization of Lipofection Protocols for CRISPR/Cas9 Delivery in Porcine Zona pellucida Intact Oocytes: A Study of Coincubation Duration and Reagent Efficacy
Theriogenology,
Год журнала:
2024,
Номер
230, С. 121 - 129
Опубликована: Сен. 14, 2024
Язык: Английский
Optimizing Electroporation Parameters for Efficient Delivery of Large Molecules into Pig Zygotes Using Fluorescent Dextrans from 3 to 2000 kDA
Juan Pablo Fernández,
Paul Kielau,
Petra Hassel
и другие.
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2024,
Номер
unknown
Опубликована: Авг. 26, 2024
Abstract
Electroporation
has
revolutionized
gene
transfer
and
editing,
enabling
efficient
delivery
of
molecules
into
embryos,
with
significant
implications
for
developmental
biology
biomedical
research.
This
study
aimed
to
optimize
electroporation
parameters
enhancing
the
large
pig
zygotes.
We
investigated
effects
fluorescence-coupled
dextran
reporters
(FDs)
sizes
ranging
from
3
2000
kiloDalton
(kDA)
along
impact
poring
polarity
settings
during
electroporation,
on
molecule
permeability.
Additionally,
we
assessed
influence
voltage
number
pulses
kDa
FDs
examined
permeability
pre-IVF
embryos
zona
pellucida-weakened
post-IVF
this
FD.
Our
findings
highlighted
size-dependent
FD
uptake,
reversing
increasing
influx
small
(3
FDs).
The
was
not
influenced
by
increased
but
it
significantly
voltage,
reaching
its
optimum
at
40
V.
in
did
show
variation
across
different
voltages.
However,
voltages
higher
than
20
V
negatively
affected
blastocyst
development
rates.
Zona-weakening
improve
offers
valuable
insights
refining
techniques
delivering
zygotes
highlights
relevance
commercial
dextrans
as
useful
tools
exploring
dynamics
electroporated
Язык: Английский