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[Porcine VHL gene cloning and construction of VHL knockdown cloned embryos].
[von hippel-lindau disease]
Von
Hippel-
Lindau
(
VHL
)
disease
is
an
autosomal
dominant
disorder
and
its
clinical
manifestation
including
haemangioblastomas
of
the
central
nervous
system
,
renal
cell
carcinoma
,
haeochromocytomas
,
and
pancreatic
cyst
.
The
deletion
,
mutation
and
promoter
methylation
of
VHL
gene
can
cause
VHL
disease
.
Swine
is
considered
as
an
ideal
model
for
human
disease
because
of
its
physiological
and
anatomical
similarity
to
human
.
We
cloned
pig
VHL
gene
that
is
2
725
bp
in
length
.
VHL
highly
expressed
in
adrenal
gland
,
liver
,
pancreas
,
heart
and
testis
.
We
designed
5
shRNAs
and
screened
the
most
effective
interference
RNA
fragment
with
a
knockdown
efficiency
of
72
%
.
Porcine
embryonic
fibroblasts
stably
transfected
with
pGenesil-shRNA
vector
were
used
as
donor
cells
for
nuclear
transfer
and
there
was
no
significant
difference
of
embryo
development
compared
with
the
control
group
.
Moreover
,
VHL
was
efficiently
knocked-down
with
efficiency
of
71
%
in
porcine
cloned
blastocyst
,
these
results
lay
a
solid
foundation
for
constructing
the
VHL
knock-down
model
of
pig
.
