Precise studies of plant, animal and human genomes enable remarkable opportunities of obtained data application in biotechnology and medicine. However, knowing nucleotide sequences isn’t enough for understanding of particular genomic elements functional relationship and their role in phenotype formation and disease pathogenesis. In post-genomic era methods allowing genomic DNA sequences manipulation, visualization and regulation of gene expression are rapidly evolving. Though, there are few methods, that meet high standards of efficiency, safety and accessibility for a wide range of researchers. In 2011 and 2013 novel methods of genome editing appeared - this are TALEN (Transcription Activator-Like Effector Nucleases) and CRISPR …
Journal of Central Nervous System Disease,
Год журнала:
2022,
Номер
14
Опубликована: Апрель 1, 2022
Huntington's
disease
(HD)
is
an
autosomal
neurodegenerative
that
characterized
by
excessive
number
of
CAG
trinucleotide
repeats
within
the
huntingtin
gene
(
International Journal of Agronomy,
Год журнала:
2017,
Номер
2017, С. 1 - 15
Опубликована: Янв. 1, 2017
The
emergence
of
genome
manipulation
methods
promises
a
real
revolution
in
biotechnology
and
genetic
engineering.
Targeted
editing
the
genomes
living
organisms
not
only
permits
investigations
into
understanding
fundamental
basis
biological
systems
but
also
allows
addressing
wide
range
goals
towards
improving
productivity
quality
crops.
This
includes
creation
plants
with
valuable
compositional
properties
traits
that
confer
resistance
to
various
biotic
abiotic
stresses.
During
past
few
years,
several
novel
have
been
developed;
these
include
zinc
finger
nucleases
(ZFNs),
transcription
activator-like
effector
(TALENs),
clustered
regularly
interspaced
short
palindromic
repeats/Cas9
(CRISPR/Cas9).
These
exciting
new
methods,
briefly
reviewed
herein,
proved
themselves
as
effective
reliable
tools
for
improvement
plants.
Angewandte Chemie,
Год журнала:
2017,
Номер
130(11), С. 2816 - 2848
Опубликована: Май 18, 2017
Abstract
Die
Regulation
biologischer
Vorgänge
in
der
Natur
unterliegt
einer
strengen
räumlichen
und
zeitlichen
Steuerung;
eines
offensichtlichsten
Beispiele
ist
die
Embryogenese
bei
Metazoen.
Zahlreiche
chemische
Werkzeuge
wurden
Zell‐
Entwicklungsbiologie
zur
Untersuchung
zellulärer
Prozesse
genutzt.
Durch
regelbare
Steuerungsmethoden
Anwendungsmöglichkeiten
dieser
Verfahren
auch
auf
komplexe
biologische
Fragestellungen
ausgeweitet.
Licht
eignet
sich
ausgezeichnet
als
externer
Impuls,
denn
es
kann
mit
sehr
hoher
räumlicher
zeitlicher
Genauigkeit
eingesetzt
werden.
Dazu
verschiedene
optisch
regulierte
entwickelt
lebende
Systeme
angewendet.
In
diesem
Aufsatz
diskutieren
wir
aktuellen
Entwicklungen
optochemischer
Werkzeuge,
darunter
niedermolekulare
Verbindungen,
Peptide,
Proteine
Nukleinsäuren,
durch
Lichteinstrahlung
irreversibel
oder
reversibel
kontrolliert
werden
können,
insbesondere
für
Anwendungen
Zellen
Tieren.
The
CRISPR/Cas9
system
provides
a
novel
and
promising
tool
for
editing
the
HIV-1
proviral
genome.
We
designed
RNA-guided
targeting
regulatory
genes
tat
rev
with
guide
RNAs
(gRNA)
selected
from
each
gene
based
on
CRISPR
specificity
sequence
conservation
across
six
major
subtypes.
Each
gRNA
was
cloned
into
lentiCRISPRv2
before
co-transfection
to
create
lentiviral
vector
transduction
target
cells.
293
T
HeLa
cells
stably
expressing
Tat
Rev
proteins
successfully
abolished
expression
of
protein
relative
that
in
non-transduced
gRNA-absent
vector-transduced
functional
assays
showed
significantly
reduced
promoter-driven
luciferase
after
tat-CRISPR
transduction,
while
revealed
gp120
rev-CRISPR
transduction.
mutated
at
Cas9
cleavage
site
high
frequency
various
indel
mutations.
Conversely,
no
mutations
were
detected
off-target
sites
had
effect
cell
viability.
further
tested
persistently
latently
HIV-1-infected
T-cell
lines,
which
p24
levels
suppressed
even
cytokine
reactivation,
multiplexing
all
gRNAs
increased
efficiency.
Thus,
may
serve
as
favorable
means
achieve
cures.
The
generation
of
gene-edited
animals
using
the
CRISPRs/Cas9
system
is
based
on
microinjection
into
zygotes
which
inefficient,
time
consuming
and
demands
high
technical
skills.
We
report
optimization
an
electroporation
method
for
intact
rat
sgRNAs
Cas9
protein
in
combination
or
not
with
ssODNs
(~100
nt).
This
resulted
frequency
knockouts,
between
15
50%
analyzed
animals.
Importantly,
as
donor
template
precise
knock-in
mutations
25-100%
animals,
comparable
to
microinjection.
Electroporation
long
ssDNA
dsDNA
donors
successfully
used
past
did
allow
genome-edited
despite
visualization
within
zygotes.
Thus,
simultaneous
a
large
number
rapid,
simple,
efficient
variety
rats.
International Journal of Molecular Sciences,
Год журнала:
2020,
Номер
21(16), С. 5665 - 5665
Опубликована: Авг. 7, 2020
Genome
editing
is
a
relevant,
versatile,
and
preferred
tool
for
crop
improvement,
as
well
functional
genomics.
In
this
review,
we
summarize
the
advances
in
gene-editing
techniques,
such
zinc-finger
nucleases
(ZFNs),
transcription
activator-like
(TAL)
effector
(TALENs),
clustered
regularly
interspaced
short
palindromic
repeats
(CRISPR)
associated
with
Cas9
Cpf1
proteins.
These
tools
support
great
opportunities
future
development
of
plant
science
rapid
remodeling
crops.
Furthermore,
discuss
brief
history
each
provide
their
comparison
different
applications.
Among
various
genome-editing
tools,
CRISPR
has
become
most
popular;
hence,
it
discussed
greatest
detail.
helped
clarify
genomic
structure
its
role
plants:
For
example,
transcriptional
control
Cpf1,
genetic
locus
monitoring,
mechanism
promoter
activity,
alteration
detection
epigenetic
behavior
between
single-nucleotide
polymorphisms
(SNPs)
investigated
based
on
traits
related
genome-wide
studies.
The
present
review
describes
how
CRISPR/Cas9
systems
can
play
valuable
characterization
rearrangement
gene
functions,
improvement
important
field
crops
precision.
addition,
speed
strategy
gene-family
members
was
introduced
to
accelerate
applications
improvement.
this,
technology
advantage
that
particularly
holds
scientist’s
mind,
allows
genome
multiple
biological
systems.
Journal of Genetic Engineering and Biotechnology,
Год журнала:
2021,
Номер
19(1), С. 125 - 125
Опубликована: Авг. 21, 2021
Genome
of
an
organism
has
always
fascinated
life
scientists.
With
the
discovery
restriction
endonucleases,
scientists
were
able
to
make
targeted
manipulations
(knockouts)
in
any
gene
sequence
organism,
by
technique
popularly
known
as
genome
engineering.
Though
there
is
a
range
editing
tools,
but
this
era
dominated
CRISPR/Cas9
tool
due
its
ease
design
and
handling.
But,
when
it
comes
clinical
applications,
CRISPR
not
usually
preferred.
In
review,
we
will
elaborate
on
structural
functional
role
designer
nucleases
with
emphasis
TALENs
system.
We
also
present
unique
features
limitations
CRISPRs
which
makes
better
than
CRISPRs.
robust
technology
used
target
specific
DNA
modifications
organism.
programmable
endonucleases-based
manipulating
tools
such
meganucleases
(MN),
zinc-finger
(ZFNs),
transcription
activator-like
effector
(TALENs),
clustered
regularly
interspaced
short
palindromic
repeats
associated
protein
(CRISPR/Cas9),
research
field
experienced
tremendous
acceleration
giving
rise
modern
precision
specificity.
Though,
CRISPR-Cas9
platform
successfully
gained
more
attention
scientific
world,
ZFNs
are
their
own
ways.
Apart
from
high-specificity,
proven
mitochondrial
(mito-TALEN),
where
gRNA
difficult
import.
This
review
talks
about
goals
fulfilled
drawbacks
provides
significant
insights
into
pros
cons
two
most
popular
mini
suggests
that,
novel
opportunities
therapeutics
being
highly
sensitive
toward
modifications.
article,
briefly
explore
special
that
indispensable
synthetic
biology.
great
perspective
providing
true
guidance
researchers
working
trait
improvement
via
editing.
