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shRNA-SIRT1 Lentivirus Plasmid (SH1001-07)

Availability: In stock

USD$360.00
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Quick Overview

ATCGbio Life Technology Inc. provides the function-validated shRNA-SIRT1 (Sirtuin 1) lentivirus plasmid. 
The product plasmids product contains 2-tubes:
1. Negative-control shRNA lentivirus plasmid  20 µl (~50 ng/µl)
2. shRNA-SIRT1 (Sirtuin 1) lentivirus plasmid   20 µl (~50 ng/µl)

shRNA-SIRT1 Lentivirus Plasmid (SH1001-07)

Details

                           Produtct Name:         shRNA-SIRT1 lentivirus plasmid  
        Official Gene Symbol                     SIRT1  
        Gene ID:         23411  
        Official Full Name:         Sirtuin 1  
        Also known as:         SIR2L1  
        Target Species:Validation:         Human Function validated  
        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#1001-07  
        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.

SIRT1 gene encodes a member of the sirtuin family of proteins, homologs to the yeast Sir2 protein. Members of the sirtuin family are characterized by a sirtuin core domain and grouped into four classes. The functions of human sirtuins have not yet been determined; however, yeast sirtuin proteins are known to regulate epigenetic gene silencing and suppress recombination of rDNA. Studies suggest that the human sirtuins may function as intracellular regulatory proteins with mono-ADP-ribosyltransferase activity. The protein encoded by this gene is included in class I of the sirtuin family. Alternative splicing results in multiple transcript variants. SIRT1 protein is a NAD dependent deactylase which has been recognized as one of the energy sensors related with motablism syndrome.

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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