Nature Biotechnology,
Год журнала:
2022,
Номер
41(1), С. 96 - 107
Опубликована: Сен. 8, 2022
Despite
the
availability
of
Cas9
variants
with
varied
protospacer-adjacent
motif
(PAM)
compatibilities,
some
genomic
loci-especially
those
pyrimidine-rich
PAM
sequences-remain
inaccessible
by
high-activity
proteins.
Moreover,
broadening
sequence
compatibility
through
engineering
can
increase
off-target
activity.
With
directed
evolution,
we
generated
four
that
together
enable
targeting
most
sequences
in
human
genome.
Using
phage-assisted
noncontinuous
evolution
and
eVOLVER-supported
continuous
evolved
Nme2Cas9,
a
compact
variant,
into
recognize
single-nucleotide
pyrimidine-PAM
sequences.
We
developed
general
selection
strategy
requires
functional
editing
fully
specified
target
protospacers
PAMs.
applied
this
to
evolve
eNme2-T.1,
eNme2-T.2,
eNme2-C
eNme2-C.NR.
Variants
eNme2-T.1
eNme2-T.2
offer
access
N
The
advent
of
clustered
regularly
interspaced
short
palindromic
repeat
(CRISPR)
genome
editing,
coupled
with
advances
in
computing
and
imaging
capabilities,
has
initiated
a
new
era
which
genetic
diseases
individual
disease
susceptibilities
are
both
predictable
actionable.
Likewise,
genes
responsible
for
plant
traits
can
be
identified
altered
quickly,
transforming
the
pace
agricultural
research
breeding.
In
this
Review,
we
discuss
current
state
CRISPR-mediated
manipulation
human
cells,
animals,
plants
along
relevant
successes
challenges
present
roadmap
future
technology.
Annual Review of Neuroscience,
Год журнала:
2022,
Номер
45(1), С. 447 - 469
Опубликована: Апрель 20, 2022
Recombinant
adeno-associated
viruses
(AAVs)
are
commonly
used
gene
delivery
vehicles
for
neuroscience
research.
They
have
two
engineerable
features:
the
capsid
(outer
protein
shell)
and
cargo
(encapsulated
genome).
These
features
can
be
modified
to
enhance
cell
type
or
tissue
tropism
control
transgene
expression,
respectively.
Several
engineered
AAV
capsids
with
unique
tropisms
been
identified,
including
variants
enhanced
central
nervous
system
transduction,
specificity,
retrograde
transport
in
neurons.
Pairing
these
AAVs
modern
regulatory
elements
state-of-the-art
reporter,
sensor,
effector
enables
highly
specific
expression
anatomical
functional
analyses
of
brain
cells
circuits.
Here,
we
discuss
recent
advances
that
provide
a
comprehensive
(capsid
cargo)
toolkit
genetic
access
molecularly
defined
types.
Science,
Год журнала:
2022,
Номер
378(6625), С. 1227 - 1234
Опубликована: Дек. 15, 2022
Synthetic
gene
circuits
that
precisely
control
human
cell
function
could
expand
the
capabilities
of
gene-
and
cell-based
therapies.
However,
platforms
for
developing
in
primary
cells
drive
robust
functional
changes
vivo
have
compositions
suitable
clinical
use
are
lacking.
Here,
we
developed
synthetic
zinc
finger
transcription
regulators
(synZiFTRs),
which
compact
based
largely
on
human-derived
proteins.
As
a
proof
principle,
engineered
switches
allow
precise,
user-defined
over
therapeutically
relevant
genes
T
using
orthogonal,
US
Food
Drug
Administration-approved
small-molecule
inducers.
Our
can
instruct
to
sequentially
activate
multiple
cellular
programs
such
as
proliferation
antitumor
activity
synergistic
therapeutic
responses.
This
platform
should
accelerate
development
translation
diverse
types
contexts.
