Difference between revisions of "Part:BBa K1777003"
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The Fudan iGEM team 2015 designed the part which has 6 miR-21 binding sites and is flanked by β-globing intron1 splicing site. We used it as the linear miR-21 sponge to sequester miR-21, compared with the blank control groups and circRNA sponge groups. In addition, adding cyclizing device to the flanking region of this part, it can be involved into a back-splicing mechanism and become a circRNA sponge.<br> | The Fudan iGEM team 2015 designed the part which has 6 miR-21 binding sites and is flanked by β-globing intron1 splicing site. We used it as the linear miR-21 sponge to sequester miR-21, compared with the blank control groups and circRNA sponge groups. In addition, adding cyclizing device to the flanking region of this part, it can be involved into a back-splicing mechanism and become a circRNA sponge.<br> | ||
− | As the controls of blank groups, the linear miR-21 sponges have the similiar constructs with "Ouroboros"([https://parts.igem.org/Part: | + | As the controls of blank groups, the linear miR-21 sponges have the similiar constructs with "Ouroboros"([https://parts.igem.org/Part:BBa_K1777016 Part:BBa_K1777016]) and only lack inverted repeat sequences.<br> |
[[File: Overview_of_the_construct_linear_miR-21_sponge_igem_2015.png| 700 px | center]] | [[File: Overview_of_the_construct_linear_miR-21_sponge_igem_2015.png| 700 px | center]] | ||
− | '''Figure 1: Construct design.''' | + | '''Figure 1: Construct design.''' <br> |
+ | [[File: ls.jpg| 700px | center]]<br> | ||
+ | '''Figure 2: Examination of efficiency of miR-21 sponges.'''<br> | ||
+ | At an appropriate molar ratio of luciferase tools and sponges,the expression of ''Renilla'' shall increase.<br> | ||
===Proof of function=== | ===Proof of function=== |
Latest revision as of 22:52, 23 September 2015
miR-21 linear sponge
(β-globin SA)-(miR-21 binding site)6-(β-globin SD) | |
---|---|
Function | Linear miR-21 sponge |
Use in | Mammalian cells |
RFC standard | RFC 10 |
Backbone | pSB1C3 |
Submitted by | [http://2015.igem.org/Team:Fudan Fudan 2015] |
The Fudan iGEM team 2015 designed the part which has 6 miR-21 binding sites and is flanked by β-globing intron1 splicing site. We used it as the linear miR-21 sponge to sequester miR-21, compared with the blank control groups and circRNA sponge groups. In addition, adding cyclizing device to the flanking region of this part, it can be involved into a back-splicing mechanism and become a circRNA sponge.
As the controls of blank groups, the linear miR-21 sponges have the similiar constructs with "Ouroboros"(Part:BBa_K1777016) and only lack inverted repeat sequences.
Figure 1: Construct design.
Figure 2: Examination of efficiency of miR-21 sponges.
At an appropriate molar ratio of luciferase tools and sponges,the expression of Renilla shall increase.
Proof of function
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 81
Illegal XhoI site found at 452 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 423