CSL (CBF1/RBP-Jk) Luciferase Reporter Lentivirus (Notch Signaling Pathway)

Catalog #

78746

$885 *

Size: 500 µl x 2

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

The CSL (CBF1/RBP-Jk) Luciferase Reporter Lentivirus (Notch Signaling Pathway) are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce most mammalian cells, including primary and non-dividing cells. These viruses contain a firefly luciferase reporter driven by multiple copies of the CSL responsive element (CBF1/RBPJκ/Suppressor of Hairless/Lag-1) located upstream of the minimal TATA promoter. The lentiviruses also contain a puromycin selection marker (Figure 1). After transduction, Notch signaling can be monitored by measuring luciferase activity.

Figure 1. Schematic of the lenti-vector used to generate the CSL (CBF1/RBP-Jk) Luciferase Reporter Lentivirus.

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Product Data Gallery

CSL driven luciferase reporter activity in HEK293 cells transduced with CSL (CBF1/RBP-Jk) Luciferase Reporter Lentivirus and/or Notch1dE Lentivirus.

Product Info

Storage and Usage

Citations

Supplied As

Two vials (500 µl x 2) of lentivirus at a titer >10⁷ TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.

Materials Required But Not Supplied

Name	Ordering Information
HEK293 Cells	ATCC #CRL-1573
Thaw Medium 1	BPS Bioscience #60187
Notch1dE Lentivirus	BPS Bioscience #78747
96-well tissue culture, clear-bottom, white plate	Corning #3610
One-Step™ Luciferase Assay System	BPS Bioscience #60690
Luminometer

Formulation

The lentivirus particles were produced in HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations by special request, for an additional fee.

Background

The Notch signaling pathway controls cell fate decisions in vertebrate and invertebrate tissues, and it is involved in embryonic development, tissue homeostasis, and regulation of immune and angiogenic systems. Notch signaling is triggered through the binding of a transmembrane ligand, present in opposing cells, to one of the four existing Notch transmembrane receptors (Notch1/ Notch2/Notch3/Notch4). This results in proteolytic cleavage of the Notch receptor, releasing the constitutively active intracellular domain of Notch (NICD). NICD translocate to the nucleus and associates with the transcription factor CSL (CBF1/RBPJκ/Suppressor of Hairless/Lag-1) and coactivator Mastermind to turn on the transcription of Notch-responsive genes. Dysfunction of Notch signaling has severe consequences, from developmental disorders to cancer (such as T cell acute lymphoblastic leukemia, T-ALL, and urothelial bladder cancer). The use of Notch inhibitors, mainly gamma-secretase inhibitors, has shown promise in cancer therapy and in regenerating tissues. Further studies will deepen our understanding of Notch signaling and will benefit future therapeutic approaches.

References

Lu F.M., et al., 1996 Natl. Acad. Sci. USA 93(11): 5663-5667.
Kanungo J., et al., 2008 Neurochem. 106: 2236-48.
Cao L. et al., 2023 Blood Adv. 1182/bloodadvances.2023010380

Storage/Stability

Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C for up to 12 months from date of receipt. Avoid repeated freeze/thaw cycles. Titers can drop significantly with each freeze/thaw cycle.

Growth Media

For best results, the use of validated and optimized media from BPS Bioscience is highly recommended. Other preparations or formulations of media may result in suboptimal performance.

Media Required for the Proposed Assay

Thaw Medium 1 (BPS Bioscience #60187): MEM medium supplemented with 10% FBS, 1% non-essential amino acids, 1 mM Na pyruvate, 1% Penicillin/Streptomycin.

Applications

Screen for activators or inhibitors of the Notch signaling pathway in cells using luciferase activity as readout.
Generate CSL-responsive Luciferase Reporter cell pools or stable cell lines by puromycin selection.
Generate a Notch1dE/CSL Luciferase Reporter cell system when used in combination with Notch1dE Lentivirus (BPS Bioscience #78747).

Shipping Temperature

-80°C

Notes

To generate a CSL Luciferase Reporter stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin (as pre-determined from a killing curve, FAQs (bpsbioscience.com)) for antibiotic selection of transduced cells, followed by clonal selection.
The following Lentivirus Reporter Controls are available from BPS Bioscience to meet your experimental needs:

Negative Control Luciferase Lentivirus (BPS Bioscience #79578): Ready-to-transduce lentiviral particles expressing firefly luciferase under the control of a minimal promoter. This negative control is important to establish the specificity of any treatments and to determine the background reporter activity.
Renilla Luciferase Lentivirus (G418 or Puromycin) (BPS Bioscience #79565): Ready-to-transduce lentiviral particles expressing Renilla luciferase under the control of a CMV promoter. The RLuc lentivirus can serve as an internal control to overcome sample-to-sample variability when performing dual-luciferase reporter assays.
Firefly Luciferase Lentivirus (BPS Bioscience #79692-G, #79692-H, #79692-P): Ready-to-transduce lentiviral particles expressing firefly luciferase under the control of a CMV promoter. It serves as a positive control for transduction optimization studies.

Biosafety
The lentiviruses are produced with SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal and are not present in the lentivirus particle. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.

Troubleshooting Guide
Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For all further questions, please email [email protected].

Warnings

Avoid freeze/thaw cycles

Scientific Category

Cell Signaling Pathway

Regulatory Status

Research Use Only

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CSL (CBF1/RBP-Jk) Luciferase Reporter Lentivirus (Notch Signaling Pathway)

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