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shRNA-BRF1 Lentivirus Plasmid (SH1001-10)

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USD$360.00
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Quick Overview

ATCGbio Life Technology Inc. provides the function-validated shRNA-BRF1 lentivirus plasmid. 
The plasmids product contains 2-tubes:
1. Negative-control shRNA lentivirus plasmid  20 µl (~50 ng/µl)
2. shRNA-BRF1 lentivirus plasmid   20 µl (~50 ng/µl)

shRNA-BRF1 Lentivirus Plasmid (SH1001-10)

Details

          Produtct Name: shRNA-BRF1 Lentivirus Plasmid
 
      Official Gene Symbol:
BRF1
 
      Gene ID: 2972  
      Official Full Name: BRF1 homolog, subunit of RNA polymerase III transcription initiation factor IIIB (S. cerevisiae)  
      Also Known As: BRF; hBRF; BRF-1; GTF3B; TAF3C; TAF3B2; TF3B90; TAFIII90; TFIIIB90  
      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#SH1001-10  
      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.

BRF1 gene encodes one of the three subunits of the RNA polymerase III transcription factor complex. This complex plays a central role in transcription initiation by RNA polymerase III on genes encoding tRNA, 5S rRNA, and other small structural RNAs. The gene product belongs to the TF2B family. Several alternatively spliced variants encoding different isoforms, that function at different promoters transcribed by RNA polymerase III, have been identified.

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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