Current Opinion in Chemical Biology,
Journal Year:
2020,
Volume and Issue:
57, P. 75 - 81
Published: June 29, 2020
Epigenetic
editing
refers
to
the
locus-specific
targeting
of
epigenetic
enzymes
rewrite
local
landscape
an
endogenous
genomic
site,
often
with
aim
transcriptional
reprogramming.
Implementing
clustered
regularly
interspaced
short
palindromic
repeat–dCas9
greatly
accelerated
advancement
editing,
yielding
preclinical
therapeutic
successes
using
a
variety
enzymes.
Here,
we
review
current
applications
these
tools
in
mammals
and
shed
light
on
biochemical
improvements
that
facilitate
versatile
applications.
Nature Communications,
Journal Year:
2018,
Volume and Issue:
9(1)
Published: May 9, 2018
CRISPR
is
becoming
an
indispensable
tool
in
biological
research.
Once
known
as
the
bacterial
immune
system
against
invading
viruses,
programmable
capacity
of
Cas9
enzyme
now
revolutionizing
diverse
fields
medical
research,
biotechnology,
and
agriculture.
CRISPR-Cas9
no
longer
just
a
gene-editing
tool;
application
areas
catalytically
impaired
inactive
Cas9,
including
gene
regulation,
epigenetic
editing,
chromatin
engineering,
imaging,
exceed
functionality
WT
Cas9.
Here,
we
will
present
brief
history
tools
describe
wide
range
CRISPR-based
genome-targeting
tools.
We
conclude
with
future
directions
broader
impact
technologies.
Nucleic Acids Research,
Journal Year:
2020,
Volume and Issue:
48(10), P. 5684 - 5694
Published: April 8, 2020
Abstract
Studies
on
biological
functions
of
N6-methyladenosine
(m6A)
modification
in
mRNA
have
drawn
significant
attention
recent
years.
Here
we
describe
the
construction
and
characterization
a
CRISPR–Cas13b-based
tool
for
targeted
demethylation
specific
mRNA.
A
fusion
protein,
named
dm6ACRISPR,
was
created
by
linking
catalytically
inactive
Type
VI-B
Cas13
enzyme
from
Prevotella
sp.
P5–125
(dPspCas13b)
to
m6A
demethylase
AlkB
homolog
5
(ALKBH5).
dm6ACRISPR
specifically
demethylates
such
as
cytochrome
b5
form
(CYB5A)
increase
its
stability.
It
can
also
demethylate
β-catenin-encoding
CTNNB1
that
contains
multiple
sites
trigger
translation.
In
addition,
system
incurs
efficient
epitranscriptome
transcripts
with
limited
off-target
effects.
Targeted
coding
oncoproteins
epidermal
growth
factor
receptor
(EGFR)
MYC
suppress
proliferation
cancer
cells.
Together,
provide
programmable
vivo
manipulation
study
genes
their
related
functions.
Recent
work
suggests
extensive
adaptation
of
transposable
elements
(TEs)
for
host
gene
regulation.
However,
high
numbers
integrations
typical
TEs,
coupled
with
sequence
divergence
within
families,
have
made
systematic
interrogation
the
regulatory
contributions
TEs
challenging.
Here,
we
employ
CARGO,
our
recent
method
CRISPR
gRNA
multiplexing,
to
facilitate
targeting
LTR5HS,
an
ape-specific
class
HERVK
(HML-2)
LTRs
that
is
active
during
early
development
and
present
in
~700
copies
throughout
human
genome.
We
combine
CARGO
activation
or
interference
to,
respectively,
induce
silence
LTR5HS
en
masse,
demonstrate
this
system
robustly
targets
vast
majority
insertions.
Remarkably,
activation/silencing
associated
reciprocal
up-
down-regulation
hundreds
genes.
These
effects
require
presence
retroviral
sequences,
but
occur
over
long
genomic
distances,
consistent
a
pervasive
function
as
embryonic
enhancers
apes.
Bioscience Reports,
Journal Year:
2020,
Volume and Issue:
40(4)
Published: March 24, 2020
Genome
editing
technologies,
particularly
those
based
on
zinc-finger
nucleases
(ZFNs),
transcription
activator-like
effector
(TALENs),
and
CRISPR
(clustered
regularly
interspaced
short
palindromic
repeat
DNA
sequences)/Cas9
are
rapidly
progressing
into
clinical
trials.
Most
use
of
to
date
has
focused
ex
vivo
gene
cells
followed
by
their
re-introduction
back
the
patient.
The
approach
is
highly
effective
for
many
disease
states,
including
cancers
sickle
cell
disease,
but
ideally
genome
would
also
be
applied
diseases
which
require
modification
in
vivo.
However,
technologies
can
confounded
problems
such
as
off-target
editing,
inefficient
or
delivery,
stimulation
counterproductive
immune
responses.
Current
research
addressing
these
issues
may
provide
new
opportunities
space.
In
this
review,
we
examine
current
status
scientific
basis
trials
featuring
ZFNs,
TALENs,
CRISPR-based
known
limitations
humans,
developing
engineering
space
that
should
lay
groundwork
further
translation
application.
