Biology Open,
Journal Year:
2023,
Volume and Issue:
12(11)
Published: Sept. 25, 2023
Christianson
syndrome
(CS)
is
an
X-linked
disorder
resulting
from
loss-of-function
(LoF)
mutations
in
SLC9A6
encoding
the
endosomal
Na+/H+
exchanger
6
(NHE6).
CS
presents
with
developmental
delay,
seizures,
intellectual
disability,
nonverbal
status,
postnatal
microcephaly,
and
ataxia.
To
define
transcriptome
signatures
of
NHE6
LoF,
we
conducted
in-depth
RNA-sequencing
(RNA-seq)
analysis
on
a
haploid
null
cell
model.
CRIPSR/Cas9
genome
editing
introduced
multiple
LoF
into
near
human
line
Hap1.
Isogenic,
paired
parental
controls
were
also
studied.
mutant
lines
confirmed
to
have
intra-endosomal
over-acidification
as
was
seen
other
cells.
RNA-seq
performed
by
two
widely
used
pipelines:
HISAT2-StringTie-DEseq2
STAR-HTseq-DEseq2.
We
identified
1056
differentially
expressed
genes
lines,
including
associated
neurodevelopment,
synapse
function,
voltage-dependent
calcium
channels,
neuronal
signaling.
Weighted
gene
co-expression
network
then
applied
critical
module
enriched
for
governing
lysosome
function.
By
identifying
significantly
changed
expression
that
lysosomal
mechanisms
NHE6-null
cells,
our
analyses
suggest
loss
function
may
converge
implicated
lysosome-related
neurologic
disease.
Further,
this
model
will
serve
important
tool
translational
science
CS.
Cell,
Journal Year:
2024,
Volume and Issue:
187(14), P. 3585 - 3601.e22
Published: May 30, 2024
Dolichol
is
a
lipid
critical
for
N-glycosylation
as
carrier
activated
sugars
and
nascent
oligosaccharides.
It
commonly
thought
to
be
directly
produced
from
polyprenol
by
the
enzyme
SRD5A3.
Instead,
we
found
that
dolichol
synthesis
requires
three-step
detour
involving
additional
metabolites,
where
SRD5A3
catalyzes
only
second
reaction.
The
first
third
steps
are
performed
DHRSX,
whose
gene
resides
on
pseudoautosomal
regions
of
X
Y
chromosomes.
Accordingly,
report
pseudoautosomal-recessive
disease
presenting
congenital
disorder
glycosylation
in
patients
with
missense
variants
DHRSX
(DHRSX-CDG).
Of
note,
has
unique
dual
substrate
cofactor
specificity,
allowing
it
act
NAD+-dependent
dehydrogenase
NADPH-dependent
reductase
two
non-consecutive
steps.
Thus,
our
work
reveals
unexpected
complexity
terminal
biosynthesis.
Furthermore,
provide
insights
into
mechanism
which
metabolism
defects
contribute
disease.
Cell Reports Methods,
Journal Year:
2024,
Volume and Issue:
4(5), P. 100776 - 100776
Published: May 1, 2024
Continual
advancements
in
genomics
have
led
to
an
ever-widening
disparity
between
the
rate
of
discovery
genetic
variants
and
our
current
understanding
their
functions
potential
roles
disease.
Systematic
methods
for
phenotyping
DNA
are
required
effectively
translate
data
into
improved
outcomes
patients
with
diseases.
To
make
biggest
impact,
these
approaches
must
be
scalable
accurate,
faithfully
reflect
disease
biology,
define
complex
mechanisms.
We
compare
analyze
function
endogenous
context
using
genome
editing
strategies,
such
as
saturation
editing,
base
prime
editing.
discuss
how
technologies
can
linked
high-content
readouts
gain
deep
mechanistic
insights
variant
effects.
Finally,
we
highlight
key
challenges
that
need
addressed
bridge
genotype
phenotype
gap,
ultimately
improve
diagnosis
treatment
Progression
through
the
G1
phase
of
cell
cycle
is
most
highly
regulated
step
in
cellular
division.
We
employed
a
chemogenetic
approach
to
discover
novel
networks
that
regulate
progression.
This
uncovered
functional
clusters
genes
altered
sensitivity
cells
inhibitors
G1/S
transition.
Mutation
components
Polycomb
Repressor
Complex
2
rescued
proliferation
inhibition
caused
by
CDK4/6
inhibitor
palbociclib,
but
not
S
or
mitosis.
In
addition
its
core
catalytic
subunits,
mutation
PRC2.1
accessory
protein
MTF2,
PRC2.2
JARID2,
rendered
resistant
palbociclib
treatment.
found
(MTF2),
(JARID2),
was
critical
for
promoting
H3K27me3
deposition
at
CpG
islands
genome-wide
and
promoters.
included
promoter
cyclins
CCND1
CCND2,
loss
MTF2
lead
upregulation
both
CCND2.
Our
results
demonstrate
role
PRC2.1,
PRC2.2,
antagonizing
progression
diversity
linages,
including
chronic
myeloid
leukemia
(CML),
breast
cancer,
immortalized
lines.
Repression
of
retrotransposition
is
crucial
for
the
successful
fitness
a
mammalian
organism.
The
domesticated
transposon
protein
L1TD1,
derived
from
LINE-1
(L1)
ORF1p,
an
RNA-binding
that
expressed
only
in
some
cancers
and
early
embryogenesis.
In
human
embryonic
stem
cells,
it
found
to
be
essential
maintaining
pluripotency.
cancer,
L1TD1
expression
highly
correlative
with
malignancy
progression
as
such
considered
potential
prognostic
factor
tumors.
However,
its
molecular
role
cancer
remains
largely
unknown.
Our
findings
reveal
DNA
hypomethylation
induces
HAP1
tumor
cells.
depletion
significantly
modulates
both
proteome
transcriptome
thereby
reduces
cell
viability.
Notably,
associates
L1
transcripts
interacts
ORF1p
protein,
facilitating
retrotransposition.
data
suggest
collaborates
ancestral
RNA
chaperone,
ensuring
efficient
retrotransposons,
rather
than
directly
impacting
abundance
targets.
this
way,
might
have
important
not
during
development
but
also
tumorigenesis.
Repression
of
retrotransposition
is
crucial
for
the
successful
fitness
a
mammalian
organism.
The
domesticated
transposon
protein
L1TD1,
derived
from
LINE-1
(L1)
ORF1p,
an
RNA-binding
that
expressed
only
in
some
cancers
and
early
embryogenesis.
In
human
embryonic
stem
cells,
it
found
to
be
essential
maintaining
pluripotency.
cancer,
L1TD1
expression
highly
correlative
with
malignancy
progression
as
such
considered
potential
prognostic
factor
tumors.
However,
its
molecular
role
cancer
remains
largely
unknown.
Our
findings
reveal
DNA
hypomethylation
induces
HAP1
tumor
cells.
depletion
significantly
modulates
both
proteome
transcriptome
thereby
reduces
cell
viability.
Notably,
associates
L1
transcripts
interacts
ORF1p
protein,
facilitating
retrotransposition.
data
suggest
collaborates
ancestral
RNA
chaperone,
ensuring
efficient
retrotransposons,
rather
than
directly
impacting
abundance
targets.
this
way,
might
have
important
not
during
development
but
also
tumorigenesis.
Nucleic Acids Research,
Journal Year:
2023,
Volume and Issue:
51(14), P. 7580 - 7601
Published: May 25, 2023
Abstract
The
selenocysteine
(Sec)
tRNA
(tRNA[Ser]Sec)
governs
Sec
insertion
into
selenoproteins
by
the
recoding
of
a
UGA
codon,
typically
used
as
stop
codon.
A
homozygous
point
mutation
(C65G)
in
human
tRNA[Ser]Sec
acceptor
arm
has
been
reported
two
independent
groups
and
was
associated
with
symptoms
such
thyroid
dysfunction
low
blood
selenium
levels;
however,
extent
altered
selenoprotein
synthesis
resulting
from
this
yet
to
be
comprehensively
investigated.
In
study,
we
CRISPR/Cas9
technology
engineer
heterozygous
mutant
cells,
which
then
compared
parental
cell
lines.
This
C65G
affected
many
aspects
integrity
activity.
Firstly,
expression
level
significantly
reduced
due
an
recruitment
RNA
polymerase
III
at
promoter.
Secondly,
strongly
altered,
but,
more
surprisingly,
it
no
longer
sensitive
supplementation.
Mass
spectrometry
analyses
revealed
isoform
unmodified
wobble
nucleotide
U34
cells
that
correlated
activities.
Overall,
study
demonstrates
pleiotropic
effect
single
on
both
phenotype
selenoproteome
expression.
Scientific Reports,
Journal Year:
2023,
Volume and Issue:
13(1)
Published: Dec. 16, 2023
Cellular
homeostasis
of
creatine
(CT),
integral
part
the
energy
buffering
and
transducing
system
connecting
intracellular
sites
ATP
production
utilization,
comprises
mechanisms
that
increase
CT,
i.e.,
biosynthesis
cellular
uptake,
CT-lowering
processes,
such
as
export
non-enzymatic
conversion
to
creatinine.
The
CT
is
controlled
by
negative
feedback
loop
via
suppression
rate-limiting
enzyme
arginine:glycine
amidinotransferase
(AGAT).
Although
regulatory
mechanism
involved
not
well
understood,
AGAT
successfully
used
in
patients
with
guanidinoacetate
methyltransferase
(GAMT)
deficiency
reduce
neurotoxic
accumulation
AGAT-mediated
supplementing
CT.
Utilizing
CT-dependent
for
upregulation
expression
may
represent
a
therapeutic
target
an
additional
syndrome,
transporter
(CrT)
defect,
which
no
effective
treatment
option
available
so
far.
We
have
CRISPR
tag
C-terminus
nanoluc
luciferase
(NLuc)
reporter
HAP1
cells.
A
biphasic
decay
AGAT-NLuc
response
increasing
extracellular
was
observed,
whereas
decrease
directly
proportional
rise
levels
approximate
IC50
1-2
mM.
generated
CrT
null
cells
stably
expressing
CrT-GFP
fusion
protein
further
demonstrated
mediated
high-affinity
(Km
9-10
µM)
dependent,
saturable
independent,
unsaturable
uptake
process.
direct
suggests
existence
sensing
enabling
dynamic
cell
changing
concentration
relevant
homeostasis.
bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2024,
Volume and Issue:
unknown
Published: April 23, 2024
Abstract
Transgene
expression
in
eHAP
cells,
a
haploid
cell
line
popularly
used
to
generate
gene
knockouts,
is
difficult
owing
its
low
transfection
efficiency
and
propensity
for
silencing
integrated
transgenes.
To
simplify
transgene
expression,
we
engineered
insulated
integrating
plasmids
that
sustain
high
levels
of
can
be
other
lines.
These
vectors
are
compatible
with
FLP-FRT
piggyBac
integration,
they
flank
gene-of
interest
bilaterally
tandem
cHS4
core
insulators,
co-express
nuclear-localized
blue
fluorescent
protein
identification
expressing
cells.
We
further
demonstrate
transgenic
cells
fused
form
heterozygous
diploid
This
method
creates
carrying
the
material
progenitors
could
also
create
defined
genotypes.
tools
expand
repertoire
experiments
performed
cultured