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shRNA-mSirt1 Lentivirus Plasmid (sirtuin 1) (SH1001-07m1)

Availability: In stock

USD$360.00
OR

Quick Overview

ATCGbio Life Technology Inc. provides the function-validated shRNA-mSirt1 (sirtuin 1) lentivirus plasmid. 
The product plasmids product contains 2-tubes:
1. Negative-control shRNA lentivirus plasmid  20 µl (~50 ng/µl)
2. shRNA-mSirt1 (sirtuin 1) lentivirus plasmid   20 µl (~50 ng/µl)

shRNA-mSirt1 Lentivirus Plasmid (sirtuin 1) (SH1001-07m1)

Details

                           Produtct Name:         shRNA-Sirt1 lentivirus plasmid  
        Official Gene Symbol:                    Sirt1  
        Gene ID:         93759  
        Official Full Name:         sirtuin 1  
        Also Known As:         Sir2; Sir2a; SIR2L1; AA673258; Sir2alpha  
        Target Species:         Mus musculus  
        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#SH1001-07m1  
        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.

Gene expression and deacetylase activity of the class III histone deacetylase Sirtuins (SIRT1-7) are the silent information regulator 2 (Sir2) family of proteins. One of the sirtuins, sirtuin 1 is widely expressed in mammalian cells and has been studied in many tissues, including liver, skeletal muscle, adipose tissue, pancreas (β-cells), brain, and endothelium. Sirtuin 1 in human is called SIRT1, and in Mus musculus it’s called Sirt1. A substantial body of evidence suggests that, like AMPK, sirtuin 1 responds to increases and decreases in nutrient availability (caloric restriction or starvation) and energy expenditure. Regulation of sirtuin1 has been attributed to changes in NAD+ abundance and the NAD+/NADH ratio, the concentration of nicotinamide (NAM), an end product of the deacetylase reaction and a sirtuin 1 inhibitor, and the activity of NAM phosphoribosyltransferase (Nampt; visfatin), which catalyzes the reconversion of NAM to NAD+. In addition, there appears to be genetic regulation of sirtuin 1 by the action of forkhead box-containing protein and p53 on the sirtuin 1 promoter as well as regulation by various other factors such as DCB1, AROS and SENP1. Sirtuin 1  has  many common target molecules including PGC1a, eNOS and FOXO, and regulates their biological actions.

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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