Part:BBa_K3730013

3UTR with miRNA combining sequence to induce degradation of the protein synthesis sequence

3' UTR: The 3'UTR part of the protein synthesis sequence can be bound by miRNA (has-miR-22, has-miR-199a, has-miR-195) and induce degradation of the protein synthesis sequence at its front end, thus realizing the directional regulation of a protein synthesis sequence by miRNA.

Our design to realize specific replication of viruses is based on the difference of miRNA between HCC cell (HEPG2 cell line) and normal cell (HEK293T cell line).
In the following figures, 3ue stands for 3UTR-empty, which is the plasmid without 3UTR sequences, as the control group.

Fluorescence intensity of cells after transfection with 3u and 3ue.

Besides, we also designed plasmids called pRNAiU6.2-22, pRNAiU6.2-195, pRNAiU6.2-199a to increase the quantity of these miRNA in HepG2 as described previously. That was how we could know if the effect of 3UTR was actually caused by miRNA or other variables. Thus, we transfected 3u and one of these plasmids above into HepG2. The group called 3u+u6 was the controlled group, and u6 referred to pRNAiU6 without extra part.

Comparatively, the expressions of eGFP were all significantly decreased in HepG2 when these miRNA were replenished in cells.

The suicide switch in E1A gene response to different types of miRNA.

Type 5 Adenovirus E1A gene + designed 3'UTR: BBa_K3730014

For full results and descriptions please turn to the result page of our wiki.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 215
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 215
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 215
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 215
  
• 1000
  COMPATIBLE WITH RFC[1000]