siRNA sequences, target sites in human, mouse, and rat mRNA of BACE1 and silencing activity of used siRNA- and shRNA-coded plasmids (presented as % of BACE1 gene expression).
Target gene (nt numbers)
% of BACE-GFP expression in DFA by siRNA (5/1/0.1 nM)*
% of BACE-GFP expression in DFA by shRNA in plasmid*#
% of BACE1 expression in SH-SY5Y (human) cells by siRNA (RT-PCR/western blot)*&
% of BACE1 expression in HEK293 (human)/M15 mice cells by shRNA in pcPURhU6@ (RT-PCR)
Sequence:
No
-sense strand-
-antisense strand-
si-0
-AAUCAGAUUGAACCUUCAUTT-
non-silencing control siRNA
100
100
86
100
-TTUUAGUCUAACUUGGAAGUA-
si-1
-UACAGGCAGCAGUAACUUUTT-
h (733–753)m (-)r (-)
27/31/58
40 (pSilencer)
nd
nd
-TTAUGUCCGUCGUCAUUGAAA-
si-2
-AGACGCUCAACAUCCUGGUTT-
h (710–730) m (-)r (-)
6/22/76
nd
61/60
nd
-TTUCUGCGAGUUGUAGGACCA-
si-3
-GAAUCAGACAAGUUCUUCAUC-
h (946–966)m (-)r (-)
0.0/1.1/16
5 (pSilencer)
55/61
nd
-GACUUAGUCUGUUCAAGAAGU-
si-4
-AAUCAGACAAGUUCUUCAUTT-
h (947-967)m (-)r (-)
0.0/1.7/15
11 (pSilencer)
34/61
nd
- TTUUAGUCUGUUCAAGAAGUA-
si-5a
-AUCAGACAAGUUCUUCAUCAA-
h (948–968) m (-)r (-)
28/35/60
12 (pcPURhU6)
nd
32/92
-GAUAGUCUGUUCAAGAAGUAG-
si-5b
-AUCGGACAAGUUCUUCAUCAA-
h (-)m (920–940)r (920–940)
nd
nd
nd
52/30
-GAUAGCCUGUUCAAGAAGUAG-
si-6
-GACUGUGGCUACAACAUUCCA-
h (1777–1797)m (1751–1771)r (1751–1771)
1.3/8/45
20 (pcPURhU6)
nd
51/71
-UUCUGACACCGAUGUUGUAAG-
*: GFP/RFP relative fluorescence of the control, HeLa cells transfected with control nonsilencing siRNA (si-0) or pSilencer-Neg plasmid/Lipofectamine 2000 were taken as a reference nonsilenced value of 100%;#: plasmid DNA 30 ng/well (96-well plate); &: 100 nM siRNA;@: plasmid DNA 2 g/well (6-well plate); Nd: not determined.
