A Novel Method for In Vivo Gene Editing in the Brain of Guppies Using Unique Nanoparticles as Delivery Vehicles
Abstract
Background
Uncovering
the
genomic
basis
of
traits
has
advanced
rapidly
in
evolutionary
biology
and
neuroscience,
largely
through
phenotypic
with
adaptive
value
been
advancing
biology,
behavior,
due
to
research
using
non-traditional
model
systems.
Direct
gene
editing
adult
brain
represents
a
crucial
next
step
linking
genotype
phenotype,
avoiding
confounding
effects
that
arise
from
modifications
during
development.
However,
implementing
these
technologies
beyond
traditional
laboratory
models
remains
challenging
delivery
limitations.
Methods
We
developed
an
intracranial
microinjection
protocol
for
guppies
(Poecilia
reticulata )
deliver
gene-editing
elements
cells.
designed
magnetic
nanoparticles
functionalized
novel
translocating
agent,
non-viral
carrier
capable
transporting
linearized
nucleic
acids
across
cellular
nuclear
membranes.
comprehensively
assessed
nanoparticle
uptake,
colocalization,
potential
health
impacts
histological
analysis,
liver
enzyme
activity
assays,
behavioral
assessments.
Results
Our
successfully
entered
cells
colocalized
nuclei
at
rates
exceeding
50%
after
two
weeks,
demonstrating
their
efficient
in
vivo
editing.
Health
assessments
showed
no
significant
cell
death
(>
80%
viability),
toxicity
(normal
ALT,
AST,
ALP
levels),
alterations
individual
social
behaviors,
confirming
nanoparticles’
biocompatibility
systemic
safety.
Conclusions
results,
combined
previous
vitro
work
our
are
effective
system
editing,
show
they
can
be
used
safe
interventions
P.
reticulata .
This
overcomes
major
technical
barrier
acid-carrying
vehicle
brain.
approach
provides
versatile
platform
studying
genetic
mechanisms
underlying
behavior
small
freshwater
fish
while
helping
overcome
limitations
conducting
functional
studies
on
non-model
organisms.
Research Square (Research Square), Год журнала: 2025, Номер unknown
Опубликована: Апрель 1, 2025
Язык: Английский
