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Construct stable reporter cell lines
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Monitor activity of transcriptional
factors naturally—within the cell nucleus
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Study signaling pathways with flow
cytometry (FACS)
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Use with nearly all cell types,
including primary and non-dividing mammalian cells
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Choose from HIV or FIV-based
lentivectors
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Clone your own TREs, or have us clone
them for you
 |
|
Fig. 1.
Activity of the vectors depends on promoter sequence and presence of a
specific transcriptional factor. |
SBI’s
PathNet™ Transcriptional
Reporter Lentivectors take advantage of novel technology for the
detection of the activation of transcriptional factors (TFs) in a natural
environment (nuclei) based on a lentivector reporter. Following transduction
of the lentivector reporter construct into mammalian cells, the construct
becomes stably integrated into the cell genome. After integration into the
genome, the reporter gene is expressed if the transcriptional factor is
present in this cell type.
Choose between three different
reporters (dscopGFP, Luciferase, and b-Galactosidase) under the control of
the minimum (m) CMV promoter. mCMV is not active without a TRE specific to a
transcriptional factor (Fig. 1). Vectors with mCMV can be used as negative
controls or as vectors for cloning any TRE of interest. After the
introduction of a specific TRE upstream of mCMV, this construct becomes a
fully active promoter dependent on the activity of the specific
transcriptional factor (TF) (Fig. 1). Vectors contain several repeats of
specific transcriptional response elements (TREs) to ensure robust
expression of the reporter gene. PathNet™ Reporter Vectors can be packaged into pseudoviral
particles using SBI's pPACK plasmid mix.
An example of functional testing of
the p53 transcriptional reporter vector is presented in the FACS analysis
below:
Fig. 2. Analysis of p53 activity in HeLa cells
transduced with different TR constructs.
A – pTRH1-mCMV-dscGFP (negative control)
B – pTRH1-CMV-dscGFP (positive control)
C – pTRH1-p53-dscGFP (basal level of p53)
D – pTRH1-p53-dscGFP (p53 was activated by PRES-229 treatment)
Product
Information:
See a
list of available reporter
vectors.
To see the sequences of available transcriptional
factor recognition elements, click here.
User Manual
Price list
TRE Library
Custom TRE Cloning Services Available
By request, we can make specific constructs of your
choice packaged in pseudoviral particles or provided as a plasmid. Click
here for more information
about our custom services.
Clone your own Transcriptional Response Element
If you wish to clone your own TRE sequences, we
also offer cloning kits.
| |
PathNet™ Transcriptional Reporter
Vector Cloning Kit |
| |
pTRF1-mCMV-dscGFP: Cat. # TR100A-1 |
| |
pTRH1-mCMV-dscGFP: Cat. # TR500A-1 |
| |
Size: 20 Reactions |
| |
Product Components: |
| |
| 50
µl |
pTRF1 or pTRH1 Vector (uncut), 200 ng/µl |
| 10
µl |
c-Fos TRE
Ligation Control, 100 nM |
|
50 µl
|
Forward (cPPT)
Primer, 10 µM |
|
50 µl
|
Reverse2 (CMV)
Primer, 10 µM |
|
|
