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shRNA-CEBPA (CCAAT/enhancer binding protein, alpha) Lentivirus Plasmid (SH1001-11A)

Availability: In stock

USD$360.00
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Quick Overview

ATCGbio Life Technology Inc. provides the function-validated shRNA-CEBPA (CCAAT/enhancer binding protein, alpha) lentivirus plasmid. 
The plasmids product contains 2-tubes:
1. Negative-control shRNA lentivirus plasmid  20 µl (~50 ng/µl)
2. shRNA-CEBPA (CCAAT/enhancer binding protein, alpha) lentivirus plasmid   20 µl (~50 ng/µl)

shRNA-CEBPA (CCAAT/enhancer binding protein, alpha) Lentivirus Plasmid (SH1001-11A)

Details

                           Produtct Name:         shRNA-CEBPA Lentivirus Plasmid
 
        Official Gene Symbol:
                   CEBPA
 
        Gene ID:         1050  
        Official Full Name:         CCAAT/enhancer binding protein (C/EBP), alpha  
        Also Known As:         CEBP; C/EBP-alpha  
        Target Species:         Human
 
        Product Validation:         shRNA Function validated  
        Technique Information:         Instruction PDF| Web Page|MSDS  
        Product Size:         20µl (~50ng/µl), store at 4°C  
        Product Category:         RNAi, Cat#SH1001-11A  
        Price (USD):         $350.00  
                     

Lentivirus plasmids will be shipped in 0.5 ml tube, and users are free to amplify. Upon receiving the tubes, keep it at 4 °C till amplification.

The protein CEBP-alpha encoded by CEBPA intronless gene is a bZIP transcription factor which can bind as a homodimer to certain promoters and enhancers. It can also form heterodimers with the related proteins CEBP-beta and CEBP-gamma. The encoded protein has been shown to bind to the promoter and modulate the expression of the gene encoding leptin, a protein that plays an important role in body weight homeostasis. Also, the encoded protein can interact with CDK2 and CDK4, thereby inhibiting these kinases and causing growth arrest in cultured cells.

The following figure is the results of experiments to prove our shRNA performance

(One shRNA sequence was selected for one target in each design)

  
         shRNA-lentivirus-performance                   Non-target   Gene-specific
               shRNA           shRNA
• shRNA Lentivirus were created to target 10 transcription factors expressed in HepG2 cells.
• 64 hrs after infection without antibiotic selection, the target gene mRNAs were down to 28.8% in average of 10 shRNA target (p<0.0001 by paired t-test). 9 out of 10 design shRNA were able to suppress more than 70%, and only one out of 10 shRNA suppressed more modestly about 60%.
• Suppression will be greater, when the cells would be cultured longer time with antibiotics selection.
      

Ten transcription factors expressed in HepG2 cells were selected for this experiment. The lentivirus plasmids expressing shRNA were made for each target gene using our propriety design scheme (One shRNA sequence was selected for one target in each design). Lentivirus was created in 12-well plates using our Lentivirus Production kit, which produces a high titer stable lentivirus solution in 48hrs. HepG2 cells were grown in 96-well palate and infected with each virus including negative control shRNA by adding 50µl crude virus solution into HepG2 cells cultured with100µl culture medium for overnight(16hrs.). The cells were incubated further 48hrs., and harvested for Direct RT real-time PCR.  As shown in the right figure, 64 hrs. after infection, target gene mRNAs were down to 28.8% in average of 10 shRNA target. 9 out of 10 design shRNA were able to suppress more than 70%, and only one out of 10 shRNA suppressed more modestly about 60%.

References 

 1. Y. Ido, et al., PLoS ONE, 2012 Apr 07( 4): e35092
 2. Lan F, et al., J Biol Chem. 2008 Oct 10;283(41):27628-35.

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