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The design and optimization of RNA trans-splicing molecules for skin cancer therapy.
[dystrophic epidermolysis bullosa]
Targeting
tumor
marker
genes
by
RNA
trans-splicing
is
a
promising
means
to
induce
tumor
cell-
specific
death
.
Using
a
screening
system
we
designed
RNA
trans-splicing
molecules
(
RTM
)
specifically
binding
the
pre-m
RNA
of
SLCO
1
B
3
,
a
marker
gene
in
epidermolysis
bullosa
associated
squamous
cell
carcinoma
(
EB-SCC
)
.
Specific
trans-splicing
,
results
in
the
fusion
of
the
endogenous
target
mRNA
of
SLCO
1
B
3
and
the
coding
sequence
of
the
suicide
gene
,
provided
by
the
RTM
.
SLCO
1
B
3
-
specific
RTMs
containing
HSV-
tk
were
analyzed
regarding
their
trans-splicing
potential
in
a
heterologous
context
using
a
SLCO
1
B
3
expressing
minigene
(
SLCO
1
B
3
-
MG
)
.
Expression
of
the
chimeric
SLCO
1
B
3
-
tk
was
detected
by
semi-quantitative
RT-PCR
and
Western
blot
analysis
.
Cell
viability
and
apoptosis
assays
confirmed
that
the
RTMs
induced
suicide
gene
-mediated
apoptosis
in
SLCO
1
B
3
-
MG
expressing
cells
.
The
lead
RTM
also
showed
its
potential
to
facilitate
a
trans-splicing
reaction
into
the
endogenous
SLCO
1
B
3
pre-m
RNA
in
EB-SCC
cells
resulting
in
tk
-mediated
apoptosis
.
We
assume
that
the
pre-selection
of
RTMs
by
our
inducible
cell-death
system
accelerates
the
design
of
optimal
RTMs
capable
to
induce
tumor
specific
cell
death
in
skin
cancer
cells
.
Diseases
Validation
Diseases presenting
"specific cell death in skin"
symptom
dystrophic epidermolysis bullosa
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