Folie 1

hordeprobableΒιοτεχνολογία

4 Οκτ 2013 (πριν από 3 χρόνια και 6 μήνες)

60 εμφανίσεις

A scenario for a genetically controlled
fission of artificial vesicles

E. Boenzli
1
, M. Hadorn
1
, M. G. Jørgensen
2
, P. Eggenberger Hotz
3
, M. M. Hanczyc
1
, T. Yomo
4

(1) Center for Fundamental Living Technology (
FLinT
), Department of Physics and Chemistry, SDU, Denmark

(2) Department of Biochemistry and Molecular Biology (BMB), SDU, Denmark

(3) The
Mærsk

Mc
-
Kinney
Møller

Institute, SDU, Denmark

(4) Department of Bioinformatics Engineering, Osaka University, Japan

References


(1)

Noireaux

V., Maeda Y.T., and
Libchaber

A. Development of an artificial cell, from self
-
organization to computation and self
-
reproduction
.

2011. Proc
Natl

Acad

Sci

U S A. 108(9): p. 3473
-
80.

(2)

White J.M. Membrane fusion
.

1992. Science. 258(5084): p. 917
-
24.

(3)
Nomura F.,
Inaba

T., Ishikawa S., Nagata M., Takahashi S.,
Hotani

H., and
Takiguchi

K. Microscopic observations reveal that
fusogenic

peptides
induce liposome shrinkage prior to membrane fusion
.

2004. Proc
Natl

Acad

Sci

U S A. 101(10): p. 3420
-
5.

(4)
http://www.iiis2010.org/iceme/website/default.asp?vc=32

(5)
Pautot

S.,
Frisken

B.J., and
Weitz

D.A. Engineering asymmetric vesicles
.

2003. Proc
Natl

Acad

Sci

U S A. 100(19): p. 10718
-
21.

(6)
Hadorn M. and Eggenberger
Hotz

P. DNA
-
mediated self
-
assembly of artificial vesicles
.

2010.
PLoS

One. 5(3): p. e9886.

(7)
Hadorn M. and Eggenberger
Hotz

P. Towards Personalized Drug Delivery: Preparation of an Encapsulated
Multicompartment

System. in 3rd
International Joint Conference on Biomedical Engineering Systems and Technologies (BIOSTEC). 2010. Valencia, Spain.





Acknowledgements


We

thank

Steen

Rasmussen

(
FLinT
,

SDU),

Tommy

Andersen

(
FLinT
,

SDU),

Ann

Zahle

Andersen

(BMB,

SDU),

Martin

R
ø
ssel

Larsen

(BMB,

SDU),

and

Davide

De

Lucrezia

(
Explora

Srl
,

Venice)

for

their

helpful

contribution

and

support
.


Introduction


The

creation

of

artificial

cells

that

bridge

the

living

and

non
-
living

world

represents

one

of

the

main

scientific

goals

of

the

21
st

century
.

A

major

task

in

creating

self
-
replicating

artificial

cells

is

controlling

the

expression

of

a

vast

number

of

proteins

as

a

result

of

the

genetic

program

of

the

cell

as

well

as

the

nongenetic

molecular

self
-
organization

processes
.

Consequently,

internally

controlled

vesicle

division

represents

one

of

the

major

challenges

in

the

creation

of

artificial

cells
1
.


Proteins

on

the

surface

of

enveloped

viruses

induce

the

fusion

of

their

own

membrane

with

the

host

cell

membrane

to

release

their

genome

into

the

cell
2
.

Using

an

artificial

system

to

analyze

membrane

fusion

processes,

it

was

shown

that

a

small

part

of

the

viral

envelope

protein

itself

has

the

potential

to

induce

fusion

of

liposomes

if

added

externally
3
.


Scenario


In

the

present

work,

we

developed

a

scenario

how

a

genetically

controlled

fission

of

vesicles

may

be

achieved

by

the

synthesis

of

small

parts

of

the

viral

envelope

protein

within

artificial

vesicles
.

The

integration

of

the

viral

proteins

to

the

inner

leaflet

of

vesicles

(Fig
.

1
)

may

induce

fission
.

The

same

mechanism

in

nature

used

to

induce

fusion

of

compartments

may

here

be

used

here

to

induce

division

of

vesicles
.

Principle of vesicle fission induced by short peptides (red/green).

Procedure


In

the

first

step,

the

viral

proteins

are

produced

externally

by

processes

of

the

organic

chemistry
.

In

the

second

step,

they

are

produced

internally

by

a

cell
-
free

transcription/

translation

system

enclosed

in

the

vesicles

(Fig
.

2
)
.


Due

to

the

association

of

the

viral

proteins

with

the

inner

leaflet

of

the

vesicles,

the

membrane

may

become

destabilized
.

A

subsequent

increase

in

the

osmotic

pressure

in

the

bulk

solution

results

in

deflated

vesicles

and

in

repeated

contacts

of

the

inner

membrane

surfaces
.

This

contact,

in

combination

with

the

destabilization

of

the

inner

leaflet,

may

lead

to

a

fusion

of

the

opposing

membrane

areas,

and

as

a

consequence,

to

fission

of

the

vesicles

(Fig
.

2
)
.

Vesicles

represent

an

ideal

tool

to

investigate

enclosed

systems

hosting

ongoing

biochemical

processes
.

Cell
-
free

transcription/translation

systems

provide

the

minimal

machinery

to

synthesize

proteins

in

vitro
4

(red/green)
.

The

vesicle

preparation

protocol,

i
.
e
.

the

water
-
in
-
oil

emulsion

transfer

method
5
,

offers

perfect

control

over

the

composition

of

both

the

bulk

solution

and

the

intravesicular

solution
.

The

versatility

of

this

vesicle

preparation

method

was

already

increased

by

collaborators

of

this

project
6
,

7
.

Figure

1

Figure

2