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shRNA-PRKAA1 Lentivirus Plasmid (SH1001-05)

Availability: In stock

USD$360.00
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Quick Overview

ATCGbio Life Technology Inc. provides the function-validated shRNA-PRKAA1 (AMPKa1: AMP-activated, alpha 1 catalytic subunit) lentivirus plasmid. 
The plasmids product contains 2-tubes:
1. Negative-control shRNA lentivirus plasmid  20 µl (~50 ng/µl)
2. shRNA-PRKAA1 (AMPKa1: AMP-activated, alpha 1 catalytic subunit) lentivirus plasmid   20 µl (~50 ng/µl)

shRNA-PRKAA1 Lentivirus Plasmid (SH1001-05)

Details

                           Produtct Name:         shRNA-PRKAA1 Lentivirus Plasmid
 
        Official Gene Symbol:
                   PRKAA1   
        Gene ID:         5562  
        Official Full Name:         AMP-activated, alpha 1 catalytic subunit  
        Also Known As:         AMPK; AMPKa1  
        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#1001-05  
        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 encoded by PRKAA1/AMPKa1 gene belongs to the ser/thr protein kinase family. It is the catalytic subunit of the 5'-prime-AMP-activated protein kinase (AMPK). AMPK is a cellular energy sensor conserved in all eukaryotic cells. The kinase activity of AMPK is activated by the stimuli that increase the cellular AMP/ATP ratio. AMPK regulates the activities of a number of key metabolic enzymes through phosphorylation. It protects cells from stresses that cause ATP depletion by switching off ATP-consuming biosynthetic pathways. Alternatively spliced transcript variants encoding distinct isoforms have been observed. The most well-known upstream kinases of AMPK are LKB1/STK11 and CamKK. Recently it has been suggested that SIRT1 facilitates LKB1/STK11-AMPK pathway by deacetylating Lysin48 of LKB1/STK11 in NAD dependent and NAD independent manner.

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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