PromoterTest Assay Kit (Promoter Construct Panel with 8 constructs), packaged lentivector Add to Cart

Cat#: PROMTEST-GVO-CT
Quantity: 1 kit (for 10 cell lines)
Price: 1045 €
Supplier: Cellecta
Shipping: Dry Ice
Please note: Genetically modified organism (GVO), see General Terms.
User Manual  

• Find the best promoter to drive expression of your gene in your cell model
• Efficiently test 8 different cDNA promoters in any mammalian cell system
• Select the optimal Cas9 construct when screening with CRISPR knockout, CRISPRi or CRISPRa systems

The PromoterTest™ Assay Kit enables you to identify a promoter optimally suited to express a cDNA of interest in any mammalian cell system. With Cellecta’s PromoterTest Kit you can assess relative promoter strength and identify an optimal promoter for any application requiring robust expression of a gene of interest.

RNA polymerase II promoters that drive cDNA expression show a wide range of variation across different cells and different culture conditions. Using the PromoterTest Assay you have a convenient approach to screen eight different promoter configurations to rapidly identify the one that is most effective for your cell line of interest.


Image
(A) Relative expression levels of 8 promoters with in different cell lines: CMV, CMV with enhancer (E/CMV), EFS (EF1-alpha gene, short promoter), EFS with enhancer (E/EFS), Rous Sarcoma Virus (RSV), RSV with Enhancer (E/RSV), mouse CMV (mCMV), Spleen focus forming virus (SSFV). Expression levels were assessed based on GFP fluorescence and normalized against the average expression for all constructs in a cell line.

(B) Expression levels of CMV, mouse CMV, and SSFV: in blood cell lines, such as SU-DHL-10, DOHH2, and Jurkat, SSFV can be much stronger than other promoters.


Utility for CRISPR-Cas9 studies
For CRISPR pooled screening with sgRNA libraries strong expression of the Cas9 protein can be important. This protein is required for the nuclease activity of the standard CRISPR knockout system and for regulation of gene repression or activation in the CRISPRi and CRISPRa systems. For this reason, Cellecta offers a range of Cas9 constructs with various promoters. The PromoterTest Kit provides a convenient way to check the responsiveness of the various promoter options before selecting a specific Cas9 construct for your cell system.

The PromoterTest Assay comes with eight lentiviral constructs pre-packaged as ready-to-transduce lentiviral particles. Each lentiviral vector carries the same green fluorescent protein (GFP) under the control of a different promoter. To use the kit, you simply need to transduce the GFP-expressing lentiviral constructs into your cell line and compare the GFP intensities from each different construct.


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