To optimize the experimental parameters of DNA vector-based siRNA constructs, we have designed positive controls , which can knock down firefly luciferase and Renilla luciferase . These can be used for initial test experiments.
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The H1.2 promoter is an engineered inducible promoter** containing a tetracycline operator (TetO1). The tetracycline operator itself has no effect on expression. When the tetracycline repressor (TetR) is present, it effectively binds the TetO1 and blocks the transcription. In the presence of tetracycline or doxycycline, the inducer binds TetR, causing the TetR protein to release the TetO1 site, derepressing transcription.
pRNATin-H1.2 is designed for mammalian transfection. Each vector carries a neomycin or hygromycin resistance gene, which can be used to establish stable cell lines, and a coral GFP marker under CMV promoter* control, which can be used to track transfection efficiency. Adenoviral, retroviral, and lentiviral expression vectors are also available for this inducible system.
Cat. No.
Vector
Promoter
Resistance
Marker
Map
Price
Selected
SD1223
H1
Neo
cGFP
$350
SD1224
H1
Hygro
cGFP
$350
SD1220
H1
Neo
-
$350
SD1229
H1
Kanamycin
cGFP
$350
SD1234
H1
Neomycin
-
$350
SD1254
H1 (inducible)
Hygro
cGFP
$350
SD1260
H1 (inducible)
Neo
cGFP
$350
We have designed a positive control for this inducible expression vector, SD1523 (pRNATin-H1.2/Neo/siFLuc) , that can knock down firefly luciferase.
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Adenoviral siRNA Shuttle Vectors:
These adenoviral shuttle vectors are compatible with the
Clontech Adeno-XTM Expression System .
Each uses either a U6 or H1 promoter to drive siRNA
expression.
Cat.
No.
Vector
Promoter
Resistance
Marker
Map
Price
Selected
SD1205
U6
Neo
-
$225
SD1206
H1
Neo
-
$225
SD1216
H1
Neo
cGFP
$350
SD1234
H1 (inducible)
Neo
-
$350
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Adenoviral siRNA Vectors:
The following GenScript adenoviral vectors are compatible
with the Stratagene AdEasyTM Adenoviral Vector
System . Each uses either an H1 or inducible H1 promoter to
drive siRNA expression. The pRNAT vectors carry a GFP marker
(coral GFP or cGFP) under CMV promoter*
control, which can be used to track transfection
efficiency.
Cat.
No.
Vector
Promoter
Resistance
Marker
Map
Price
Selected
SD1209
H1
Kanamycin
-
$225
SD1219
H1
Kanamycin
cGFP
$350
SD1229
H1 (inducible)
Kanamycin
cGFP
$350
Lentiviral siRNA Vectors:
GenScript Lentiviral siRNA expression vectors are
compatible with the Invitrogen ViraPowerTM Lentiviral
Expression System . They are designed for siRNA expression
in both dividing and nondividing cells. Each vector contains a
neomycin resistance gene for establishing stable cell lines
and a coral GFP gene for tracking transfection efficiency,
respectively driven by an SV40 promoter and a CMV promoter.
The vector uses a Rous sarcoma virus (RSV)
enhancer/promoter fused to HIV 5'LTR (PRSV/5'LTR) for viral
transcription and HIV 3'LTR for packaging.
Cat.
No.
Vector
Promoter
Resistance
Marker
Map
Price
Selected
SD1259
U6
Neo
cGFP
$350
SD1260
H1 (inducible)
Neo
cGFP
$350
Retroviral siRNA Vectors:
GenScript Retroviral siRNA expression vectors are
compatible with the Clontech Retro-XTM Expression
System . The siRNA insert is cloned
between the Mli and Xho sites. An H1
promoter drives siRNA expression. The vector contains a
hygromycin resistance gene under the control of a CMV
promoter*
for establishing stable cell lines. The vector uses MSV 3'LTR
for viral transcription and a CMV enhancer/promoter joined to
MSV 5'LTR (CMV/MSV 5'LTR) for packaging.
