DNA

Part:BBa_K313010:Experience

Designed by: Ryosuke Kamei, Ryo Kariyazono, Takashi Matsumoto, Takashi Hiroi, Mei Kobayashi, Yuka Yashiro, Tomoka Ao   Group: iGEM10_UT-Tokyo   (2010-10-22)


We are SYSU-CHINA 2021 and this year we use circRNA to colocalize enzymes, which are fused to RNA binding proteins(RBP). We choose MS2 coat protein and PP7 coat protein as our RBP,and their aptamer on circRNA.For contribution, we predict its 2D and 3D structure,to present a visual impression.

Applications of BBa_K313010

We are SYSU-CHINA 2021 and this year we use circRNA to colocalize enzymes, which are fused to RNA binding proteins(RBP). We choose MS2 coat protein and PP7 coat protein as our RBP,and their aptamer on circRNA.For contribution, we predict its 2D and 3D structure,to present a visual impression.

2D stucture

UNIQ8a3d6ceaae184ac0-partinfo-00000000-QINU UNIQ8a3d6ceaae184ac0-partinfo-00000001-QINU
We use RNAfold and StructureEditor to predict 2D structure of our circRNA, which contains MS2 aptamer.The results are below:

MS2 aptamer on the circRNA scaffold.png
Figure 1.2D stucture predicted by RNAfold

SYSU-CHINA circRNA-1.png
Figure 2.2D stucture predicted by StructureEditor

3D stucture

UNIQ8a3d6ceaae184ac0-partinfo-00000002-QINU UNIQ8a3d6ceaae184ac0-partinfo-00000003-QINU
We use this website:http://biophy.hust.edu.cn/new/3dRNA/create, to predict the 3D structure of our circRNA, which contains MS2 aptamer.The result lists below:


SYSU-CHINA circRNA-2.png

Figure 3. 3D stucture of MS2 aptamer predicted by Xiaolab

MS2 binding mechanism

UNIQ8a3d6ceaae184ac0-partinfo-00000004-QINU UNIQ8a3d6ceaae184ac0-partinfo-00000005-QINU


MS2 binding mechanism.png

Figure 4. MS2 binding mechanism


split EGFP

UNIQ8a3d6ceaae184ac0-partinfo-00000006-QINU UNIQ8a3d6ceaae184ac0-partinfo-00000007-QINU
We are SYSU-CHINA,and this year we use this part as MS2 coat protein's aptamer to do a split EGFP experiment,for the proof of concept.Here we found a research article, which use liner RNA with MS2 and PP7 aptamer, to do the split EGFP experiment.Below are their experimental data.


The principle of spite EGFP.png

Figure 3.the principle and experimental data of split EGFP