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Cignal™ Lenti Renilla Control (luc)

Description
The Cignal Lenti Reporters are ready-to-transduce lentiviral particles for assessing cell signaling activities in virtually any mammalian cell type. The Cignal Lenti Renilla controls (luc) are ready-to-transduce lentiviral particles constitutively expressing Renilla luciferase. This is an essential internal control for normalizing your primary reporter results. It minimizes effects of cell number, cell health, transduction efficiency and non-specific cellular responses. The Cignal Lenti Renilla control (luc) is produced from the lentiviral backbone vector containing a gene encoding Renilla luciferase driven by CMV enhancer/promoter.

Why Cignal Lenti Renilla Control (luc)?

  • Serves as an internal control for normalization
  • Increase accuracy of results
  • Enables convenient and homogenous dual-luciferase assay

For more information about the Cignal Lenti Reporters, including performance data, please visit the Cignal Lenti Reporter Assay home page.

Useful Links
Pricing and Ordering Response Element Sequence Other Lenti Reporter
User Manual White Paper Transfection Reagent
Register for Webinar  Technical Support 
Related Products
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Materials Included / Packing List:
Please check the kit components immediately after you receive this package. SABiosciences is only responsible for missing items reported within two (2) business days of receipt.

Kit Contents:
Component Specification Concentration* (total volume)
Renilla luciferase Control Reporter - 1 tube Ready-to-transduce lentivirus constitutively expressing Renilla luciferase ≥ 0.8 X 107 TU/ml (250 µl)**
Renilla luciferase Control Reporter - 8 tube Ready-to-transduce lentivirus constitutively expressing Renilla luciferase ≥ 0.8 X 107 TU/ml (2000 µl)***

* exact titer specified on Certificate of Analysis

** Supplied material provides sufficient lentivirus to transduce 2.5X106 cells at a multiplicity of infection (MOI) of 1 TU per cell (refer to Cignal Lenti Reporter User Manual). 

*** Supplied material provides sufficient lentivirus to transduce 2X107 cells at a multiplicity of infection (MOI) of 1 TU per cell (refer to Cignal Lenti Reporter User Manual).

Storage Conditions: The Cignal Lenti Reporters are shipped on dry ice. Store all tubes at -80  ºC.

Advantage of Using Cignal Lenti Renilla Control

To show the importance of Cignal™ Lenti Renilla Control (luc) comparative reporter assays were carried out with or without Cignal™ Lenti Renilla Control (luc). In the experiment, Cignal Lenti HIF-1 reporter (luc) [1.5X105 TU] transduced around 15,000 HepG2 cells (24 hours before transduction 7,500 cells were plated per well of 96-well plate) with and without Cignal Lenti Renilla control (luc) [3X104 TU]. After 42 hours of transduction, medium was changed to assay medium (Opti-MEM + 0.5% FBS + 0.1mM NEAA + 1mM Sodium pyruvate + 100 U/ml penicillin + 100 µg/ml streptomycin). After 48 hours of transduction, cells were treated with 300 µM; of CoCl2 for 18 hours. Luciferase or dual-Luciferase assay was performed, and promoter activity values are expressed as arbitrary units using a Renilla reporter for internal normalization. Experiments were done in triplicates, and the standard deviation is indicated. The utilization of Cignal Lenti Renilla control along with Cignal Lenti reporter (luc) improves the fold induction change in hypoxia signaling activity, as normalization of firefly luciferase activity with renilla luciferase activity corrects for cell death caused by the treatment.

Biosafety Features
The Cignal Lenti Reporters are ready-to-transduce, replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles. The Cignal Lentiviral particles are safe to use. It is recommended that they be treated as Risk Group Level 2 (RGL-2) organisms. Follow all published RGL-2 guidelines for handling and waste decontamination. Details on the requirements for creating a BSL-2 work environment are available in the U.S. Department of Health and Human Services publication Biosafety in Microbiological and Biomedical Laboratories, 4th edition.

The biosafety features engineered into these vectors include:

  • A deletion in the promoter/enhancer region of the U3 portion of 3'LTR ensures self-inactivation of the lentiviral construct after transduction and integration into the genomic DNA of target cells.

  • The Cignal Lentiviral vector and plasmids expressing packaging proteins contain no significant areas of homology, thereby minimizing any chance for recombination.

  • None of the HIV-1 genes (gag, pol, rev) will be expressed in transduced cells, as they are expressed from packaging plasmids lacking packaging signal. Therefore, the lentiviral particles that are generated are replication-incompetent

  • No virulence genes ( vpr, vif, vpu and nef) are present in the Cignal Lentiviral vector.

 
Feature Function
RSV-5' LTR; Hybrid Rous sarcoma Virus (RSV) enhancer/promoter-U5 long terminal repeat Permits viral packaging and reverse transcription of viral mRNA
Psi; Packaging signal Allow viral packaging
RRE; Rev response element Involved in packaging of viral transcript
cppt; Central polypurine tract Involved in nuclear translocation and integration of transduced viral genome
Renilla (Renilla luciferase) Act as an internal control for normalization
hPGK; human phosphoglycerate kinase eukaryotic promoter Permits high-level expression of the mammalian selection marker (puromycin)
PuroR; puromycin resistance gene Can be used for mammalian selection
SIN/3'LTR; 3' self inactivating long terminal repeat Modified 3'LTR that allows viral packaging but self inactivates the 5'LTR for biosafety purpose. The element also contains a polyadenylation signal for efficient transcription termination
f1 ori; f1 origin of replication Origin of DNA replication for bacteriophage f1
AmpR; ampicillin resistance gene Allows selection of the plasmid in E.coli
CMV promoter  Permits high level expression of reporter gene