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sh-LTviral-mSirt1 Lentivirus Particle (sirtuin 1) (SH3001-07m1)

Availability: In stock

USD$360.00
OR

Quick Overview

sh-LTviral-mSirt1 (cat#SH3001-07m1) is the function-validated shRNA targeted Mus musculus Sirt1 (Sirtuin 1) lentivirus particle. 
The shRNA lentivirus particle product contains 3-tubes:
a). Negative-control shRNA lentivirus particel  9.0ml, up to 0.9 x 108 TU/ml
b). sh-LTviral-mSirt1 lentivirus particle   9.0ml, up to 0.9 x 108 TU/ml
c). Lenti-Storage Solution(reconstitution) 1.0ml, shipped at room temperature, store at 4ºC.
The total price for the first order is $350.00. From the second time order, the same target and control only cost $200.00.
Please CONTACT US by fill-in your fist-time order number, date and contact information including name, email, phone and address. We will provide you a second order code.

sh-LTviral-mSirt1 Lentivirus Particle (sirtuin 1) (SH3001-07m1)

Details

                           Produtct Name:         shRNA-mSirt1 lentivirus particle  
        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:         User Manual PDF | MSDS PDF  
        Product Size:         9.0ml, store at -20°C  
        Product Category:         RNAi, Cat#SH3001-07m1  
        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.

ATCGbio Life Technology offers shRNA Lentivirus particle solutions which are recombinant shRNA expression lentivirus particle to be ready to infect directly for knocking-down gene of interest. It’s specifically designed for scientists who don’t have time or don’t prefer to make lentivirus by themselves. It’s the easiest way to knockdown gene of interest.

shRNA-Lenti particle from ATCGbio Life Technology are created base on safer 3rd. generation recombinant lentivirus vector through our new shRNA design system; and produced by our own Lentivirus Production Kit (LT1001). Virus expresses shRNA for target gene, GFP protein and hygromycine resistance gene product. It is suitable for both cultured cells and in vivo injection to small animals.  Lentivirus solution is DMEM based medium containing 2% FBS and our lentivirus precipitation/purification solution. It does not contain any chemicals used to boost lentivirus titers such as chloroquin, acetate or nicotines. The solution can be used directly to the cells growing in DMEM base medium without further precipitation/purification. There is no visible viral toxicities (e.g. change in cell morphology or/and cell detachment from culture plate) if the viral solution added up to a half amount of cell culture medium as described in table 1. Some special cells such as keratinocytes may be used to grow with special medium which is incompatible with DMEM or FBS. In such a case, it is recommended to precipitate/purify lentivirus and reconstitute it with storage solution as described in User Manual.

The highlight features of the products are:

1. Most advanced shRNA design.No need to add polybrene.Near 100% transduction (Fig.1.).
2. High infectious efficiency(Fig.2.), up to 0.9 x 108 TU/ml.
3. Co-express GFP protein and hygromycine resistance gene.
4. Just add and incubate overnight (16 hrs).
5. No cytotoxicity. Injectable to small animal without immonoreactive symptoms.
6. Unlike siRNA, transduced cells show knock-down effect in both dividing and nondividing cells.
7. Cell-line can be made in a week.
8. Suitable for both primary cells and transformed cells.

Our lentivirus particle will be shipped in propriety solution making high transduction without polybrene and unprecedented stability:

• Virus solution can be stored at -20 °C.
• Allow freezing and thawing up to 2 times.
• Be stable in room temperature at least 3-4 days.
• If necessary (the cells are incompatible with DMEM base medium), purification is easy as just centrifuge for 30min at 1500 × g.

              Table.1.

                           Size of Culture          Culture Medium         Lentiviral Solution      
  1 well of 6-well plate 1.0ml 0.5ml
  1 well of 12-well plate 0.5ml 0.25ml
  1 well of 24-well plate 0.25ml 0.125ml
       
Fig.1. HepG2 cells grown in 100µl MEM+10% FBS on the 96-well plate were infected with 50µl shRNA lentivirus solution 48hrs.
Near 100% transduction.
     
Fig.2. 64hrs after infection without antibiotic selection, the target gene mRNAs were down to 28.8% in average of 10 shRNA targets in HepG2 cells.
More details are described below.
HepG2-celld-were-infected-with-50µl-shRNA-lentivirus-solution-48hrs            
     

shRNA-lentivirus-performance                                  Non-target Gene-specific                             shRNA        shRNA

As shown in Figure 2. 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 targets (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.

References

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

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