Difference between revisions of "Part:BBa K1633003:Experience"

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===Applications of BBa_K1633003===
 
===Applications of BBa_K1633003===
  
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'''INTRODUCTION'''
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==== Usage and Biology ====
  
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This part is an artificially designed RNA strand. It serves as an element of the Team NJU-CHINA RNAi module. We use it as the siRNA medicine to downregulate the expression of Mu opioid receptor in brain tissue. We designed specific MOR siRNAs based on a free software accessible online. This tool can find the best siRNA sequences on target gene MOR to insure the maximum gene-specificity and silencing efficacy. This tool also designs the pair of oligonucleotides needed to generate short hairpin RNAs (shRNAs) in the plasmid. Then we synthesize the shRNA sequences with the help of a DNA synthesis company. We totally got four such shRNA plasmids.Although MOR siRNA-1 plasmid can efficiently knock down MOR expression, it does not show best interference efficiency and therefore serve as a backup.
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This part is a shRNA designed to target and degrade MOR mRNA. When this shRNA sequence is cut by restriction enzyme and then integrated into mammalian vector, this shRNA can play a RNAi function in mammalian cell lines. When the shRNA vector of MOR is transfected into mammalian cells, the shRNA hairpin structure is cleaved by Dicer into siRNA of MOR and loaded into the RISC. The siRNA-RISC complex targets at MOR mRNA under the guide of siRNA sequence and cleave the MOR mRNA.
  
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[[File:Example.jpg]]
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==== CHARACTERIZATION ====
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Interference efficiency of MOR siRNA-1 plasmid
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To ensure the interference efficiency, MOR siRNA-1 plasmid was,transfected into the mouse neuroblastoma cell line Neuro2A. Efficient knockdown of MOR by MOR siRNA-1 in Neuro2A cells is observed.
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[[File:NJU-China-parts-fig 2.png|400px]]
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Figure 2. Relative level of MOR mRNA in Neuro2A cell after transfection of MOR siRNA-1 plasmid.
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 14:35, 18 September 2015

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1633003

Usage and Biology

This part is a shRNA designed to target and degrade MOR mRNA. When this shRNA sequence is cut by restriction enzyme and then integrated into mammalian vector, this shRNA can play a RNAi function in mammalian cell lines. When the shRNA vector of MOR is transfected into mammalian cells, the shRNA hairpin structure is cleaved by Dicer into siRNA of MOR and loaded into the RISC. The siRNA-RISC complex targets at MOR mRNA under the guide of siRNA sequence and cleave the MOR mRNA.

CHARACTERIZATION

Interference efficiency of MOR siRNA-1 plasmid

To ensure the interference efficiency, MOR siRNA-1 plasmid was,transfected into the mouse neuroblastoma cell line Neuro2A. Efficient knockdown of MOR by MOR siRNA-1 in Neuro2A cells is observed.

NJU-China-parts-fig 2.png

Figure 2. Relative level of MOR mRNA in Neuro2A cell after transfection of MOR siRNA-1 plasmid.

User Reviews

UNIQe2ba4f61981f1a10-partinfo-00000000-QINU UNIQe2ba4f61981f1a10-partinfo-00000001-QINU