of 5
Supplementary information
Gold
N
anocluster
M
ediated
D
elivery of siRNA to
I
ntact
P
lant
C
ells for
E
fficient
Gene Knockdown
Huan Zhang
1,
2,
#
, Yuhong Cao
3
,
#
, †
, Dawei Xu
2
, Natalie S. Goh
2
,
Gozde S. Demirer
2, $
,
Stefano
Cestellos
-
Blanco
3
,
Yuan Chen
4
, Ma
r
kita P
.
Landry
2
,
5
,
6*
, Peidong Yang
3
,
7
,
8
,
9*
1.
Department of
Chemistry,
College of Chemistry and
Material
s
Science,
Jinan University, Guangzhou,
Guangdong 510632, China
2.
Department of Chemical and
B
iomolecular Engineering, Univer
sity of California, Berkeley, CA 94720,
USA
3.
Department of Materials Science and Engineering, University of California, Berkeley, CA 94720
4.
Plant Gene Expression Center, United States Department of Agriculture
-
Agricultural Research Service,
and
Department of Plant and Microbial Biology, University of California Berkeley, Albany, California
94710
5.
California Institute for Quantitative Biosciences, QB3, University of California, Berkeley, CA 94720,
USA
6.
Chan Zuckerberg BioHub, San Francisco, CA 9415
8, USA
7.
Department of Chemistry, University of California, Berkeley, CA 94720
8.
Chemical
Sciences Division,
Lawrence Berkeley National Laboratory, Berkeley,
CA
94720
9.
Kavli Energy NanoScience
Institute,
University of California, Berkeley,
CA
94720
Current address
:
CAS Key Laboratory for Biological Effects of Nanomaterials and Nanosafety, National
Center of Nanoscience and Technology, 11 Zhongguancun Beiyitiao Haidian District, Beijing, China
,
100190
$
Current address: Department of Plant Biology and Genome Center, University of California, Davis, 451
Health Sciences Drive, Davis, CA 95616, USA
*
_
Corresponding authors, Email: p_yang@berkeley.edu and landry@berkeley.edu;
#
_
H.Z. and Y.C.
c
ontributed equally to this work.
Supplementary
Ta
ble
S
1
.
Dynamic
light
scattering (DLS) size and Z
eta
potential
measurements of PEI
-
AuNCs and siRNA loaded PEI
-
AuNCs.
Name
Average size (nm)
PDI
Zeta potential (mV)
800
-
PEI
AuNCs
6.46
0.598
+
33.63
2.5
K
-
PEI
AuNCs
5.11
0.542
+
54.2
0
25
K
-
PEI
AuNCs
6.53
0.562
+
36.3
7
800
-
PEI
AuNCs+siRNA
19.58
0.353
+29.50
2.5
K
-
PEI AuNCs+siRNA
27.69
0.46
0
+36.67
25
K
-
PEI
AuNCs+siRNA
24.42
0.382
+35.7
0
Supplementary Table S
2
.
Sequences of oligonucleotides used in this study.
Name
Sequence (5’
-
3’)
Cy3
-
DNA
CTAAGGATGCGTGTA
-
Cy3
sense RNA
rGrGrUrGrArUrGrCrArArCrArUrArCrGrGrArATT
a
ntisense
RNA
rUrUrCrCrGrUrArUrGrUrUrGrCrArUrCrArCrC
m
GFP
forward
AGTGGAGAGGGTGAAGGTGATG
m
GFP
reverse
GCATTGAACACCATAAGAGAAAGTAGTG
EF1
forward
GCATTGAACACCATAAGAGAAAGTAGTG
EF1
reverse
ACGCTTGAGATCCTTAACCGCAACATTCTT
NbrbohB
forward
TTTCTCTGAGGTTTGCCAGCCACCACCTAA
NbrbohB
reverse
GCCTTCATGTTGTTGACAATGTCTTTAACA
Figure
S
1
.
TEM
image
and size distribution analysis of
siRNA
-
loaded
2.5
K
-
PEI
AuNCs.
F
igure S
2
.
Representative confocal images
of
Cy
3
-
DNA
loaded
2.
5K
-
PEI
AuNC
-
infiltrated
Nb
leaf cells
at different incubation time
points (from 20
min
ute
s to 24
h
ours
)
.
Scale Bar: 100
μ
m.
Figure
S
3
.
Original western blot image of
GFP
a
) and tubulin
b
)
extracted from
different
treated
leaves
3 days
post
-
infiltration
.
F
igure S
4
.
qPCR analysis of
ROQ1
gene 1
-
day post
infiltration
of samples into
Nb
leaf tissues
.
****p<0.0001, ***p<0.001 in one
-
way ANOVA
; Error bars indicates s.e.m (n = 3).
Figure
S
5
.
qPCR analysis of
NbrbohB
gene 1
-
day post infiltration
of samples into
Nb
leaf tissues
.
n.s.: not significant; Error
bars indicates s.e.m (n = 3).