Difference between revisions of "Team:SMMU-China/Part Collection"

Line 87: Line 87:
 
<h2 class="inner-h">Abstract</h2>
 
<h2 class="inner-h">Abstract</h2>
 
<p class="inner-text">
 
<p class="inner-text">
−
This year is the first year which SMMU-iGEM 2018 team becomes a family member of iGEM. We have designed 14 parts, including 6 basic parts. All of the parts are designed ourselves and most of them have been tested to insure their reliability.
+
This year is the first year which SMMU-iGEM 2018 team becomes a family member of iGEM.We have designed altogether 13 parts, including 5 basic parts and 8 composite parts. The purpose of these parts is to specifically express intracellular nanobody AR185 in failing cardiomyocytes conducted by AAV9 vector. Our favorite parts are coding sequnce AR185-T2A-EGFP (BBa_K2865001), heart failure inducible BNP promoter (BBa_K2865000), and composite part BBa_K2865013, which is used to package AAV9 containing our therapeutic gene AR185 driven by CMV promoter. We have tested the above parts and results suggested that AR185 was effective and there were significant differences between AR185 group and control group. The package of AAV9 virus succeeded as well. To our dismay, BNP promoter didn’t response well to HF-related factors AngⅡ and ET-1 in our vitro cell experiments, and its function remains to be investigated further. The part numbers range from BBa_K2865000 to BBa_K2865015, except for BBa_K2865007 and BBa_K2865011.
−
</p>
+
−
<p class="inner-text">
+
−
As a team focusing on real-world problem, heart faliure, we found it is hard to control the process of disease through traditional drug threopy, implanted cardiac assist devices or surgical treatments. In synthetic biology, one central route is to construct a controllable biological network which could express in the environment of heart failure. We designed a targeting device, Ca<sup>2+</sup>RTIN (Cardiomyocyte RyR2 Targeting Intra-Nanobody), to implement RyR2-specific inhibition of phosphorylation.
+
−
</p>
+
−
<p class="inner-text">
+
−
In this year, we investigated the potential of adeno-associated virus (AAV)-9-mediated cardiac expression of AR185 which is one of the isolated nanobodies from a phage display library of variable domains of camellidae heavy chain-only antibodies (VHH) to combat post-ischemic heart failure. To achieve controlled intra-nanobody release, a BNP promoter based platform were also accessed. Our results established a role of Ca<sup>2+</sup>RTIN as a promising therapeutic approach for heart failure.
+
−
</p>
+
−
<p class="inner-text">
+
−
Our members are involved in the construction of the parts, and has achieved some results. The functions of all components are tested and validated under controlled laboratory conditions and normal conditions, and data and phenomena demonstrate that they are valid. At the same time, all components are standardized, which make them a convenient and powerful tool for others.
+
 
</p>
 
</p>
 
<div style="text-align:center" class="resultimage">
 
<div style="text-align:center" class="resultimage">
−
<img src="https://static.igem.org/mediawiki/2018/a/a1/T--SMMU-China--Part_Collection_Pic_1.jpg" style="width: 60%;">
+
<img src="https://static.igem.org/mediawiki/2018/9/9a/T--SMMU-China--Part_Collection_Pic_1.png">
 
<p style="font-style: italic;text-align: center;padding: 0em 100px 1em;">
 
<p style="font-style: italic;text-align: center;padding: 0em 100px 1em;">
−
<strong>Design and Function of Ca<sup>2+</sup>RTIN</strong>
+
<strong>Design and Function of part of Left ITR-CMV-AR185-Poly(A)-Right ITR</strong>
 
</p>
 
</p>
 
</div>
 
</div>
Line 133: Line 124:
 
<td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2865001" target="_Blank">BBa_K2865001</a></td>
 
<td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2865001" target="_Blank">BBa_K2865001</a></td>
 
<td>Coding</td>
 
<td>Coding</td>
−
<td>AR185, nanobody inhibiting RyR2 phosphorylation</td>
+
<td>AR185-T2A-EGFP, nanobody inhibiting RyR2 phosphorylation</td>
 
<td>1204</td>
 
<td>1204</td>
 
</tr>
 
</tr>
Line 172: Line 163:
 
</tr>
 
</tr>
 
<tr>
 
<tr>
−
<td><i class="fa fa-heart" style="color: #F40"></i></td>
+
<td></td>
 
<td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2865009" target="_Blank">BBa_K2865009</a></td>
 
<td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2865009" target="_Blank">BBa_K2865009</a></td>
 
<td>Composite</td>
 
<td>Composite</td>
Line 193: Line 184:
 
</tr>
 
</tr>
 
<tr>
 
<tr>
−
<td></td>
+
<td><i class="fa fa-heart" style="color: #F40"></i></td>
 
<td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2865013" target="_Blank">BBa_K2865013</a></td>
 
<td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2865013" target="_Blank">BBa_K2865013</a></td>
 
<td>Composite</td>
 
<td>Composite</td>

Revision as of 01:49, 14 October 2018

Part Collection

Abstract

This year is the first year which SMMU-iGEM 2018 team becomes a family member of iGEM.We have designed altogether 13 parts, including 5 basic parts and 8 composite parts. The purpose of these parts is to specifically express intracellular nanobody AR185 in failing cardiomyocytes conducted by AAV9 vector. Our favorite parts are coding sequnce AR185-T2A-EGFP (BBa_K2865001), heart failure inducible BNP promoter (BBa_K2865000), and composite part BBa_K2865013, which is used to package AAV9 containing our therapeutic gene AR185 driven by CMV promoter. We have tested the above parts and results suggested that AR185 was effective and there were significant differences between AR185 group and control group. The package of AAV9 virus succeeded as well. To our dismay, BNP promoter didn’t response well to HF-related factors AngⅡ and ET-1 in our vitro cell experiments, and its function remains to be investigated further. The part numbers range from BBa_K2865000 to BBa_K2865015, except for BBa_K2865007 and BBa_K2865011.

Design and Function of part of Left ITR-CMV-AR185-Poly(A)-Right ITR

See More

To see their specific function, you can click the number link below to browse the specific page on parts.igem.org.

We listed Biobricks belong to Part Collection below.

Favourite Name Type Description Length
BBa_K2865000 Regulatory BNP promoter, heart failure inducible 508
BBa_K2865001 Coding AR185-T2A-EGFP, nanobody inhibiting RyR2 phosphorylation 1204
BBa_K2865002 Project [AAV9]-Left-ITR 175
BBa_K2865003 Project [AAV9]-Right-ITR 207
BBa_K2865004 Terminator SV40 poly(A) 261
BBa_K2865006 Composite CMV promoter-eGFP-poly(A) 1666
BBa_K2865008 Composite BNP promoter-eGFP-poly(A) 1520
BBa_K2865009 Composite BNP-AR185-Poly(A) 1989
BBa_K2865010 Composite CMV-AR185-Poly(A) 2135
BBa_K2865012 Composite Left ITR-BNP-AR185-Poly(A)-Right ITR 2387
BBa_K2865013 Composite Left ITR-CMV-AR185-Poly(A)-Right ITR 2533
BBa_K2865014 Composite Left ITR-CMV-eGFP-Poly(A)-Right ITR 2064
BBa_K2865015 Composite Left ITR-BNP-eGFP-Poly(A)-Right ITR 1918

References

  1. Sergeeva, I.A. and V.M. Christoffels, Regulation of expression of atrial and brain natriuretic peptide, biomarkers for heart development and disease. Biochim Biophys Acta, 2013. 1832(12): p. 2403-13.
  2. Bingham, A.J., et al., The repressor element 1-silencing transcription factor regulates heart-specific gene expression using multiple chromatin-modifying complexes. Mol Cell Biol, 2007. 27(11): p. 4082-92.
  3. Kerkela, R., et al., Key roles of endothelin-1 and p38 MAPK in the regulation of atrial stretch response. Am J Physiol Regul Integr Comp Physiol, 2011. 300(1): p. R140-9.
  4. Lu, Y.M., et al., DY-9760e inhibits endothelin-1-induced cardiomyocyte hypertrophy through inhibition of CaMKII and ERK activities. Cardiovasc Ther, 2009. 27(1): p. 17-27.
to the top