Difference between revisions of "Part:BBa K1993014"

 
(3 intermediate revisions by 2 users not shown)
Line 12: Line 12:
  
 
<html><p align="center">
 
<html><p align="center">
<img src="" style="width:400px"  ></a>
+
<img src="https://static.igem.org/mediawiki/2016/2/29/T--SYSU-MEDICINE--pSMA-eGFP.png" style="width:400px"  ></a>
  
 
</p></html>  <br>
 
</p></html>  <br>
  
<p align="center">'''_______________'''</p><br>
+
<p align="center">'''Figure 1 pSMA-eGFP'''</p><br>
  
To confirm the function of this plasmid, we administered TGF-β1 (7ng/mL). After that, α-
+
To confirm the function of this plasmid, we administered TGF-β1 (7ng/mL). After that, Promoter of α-SMA was activated and green fluorescence could be detected in MSCs (Figure 2)
SMA was expressed and green fluorescence could be detected in MSCs (Figure 2)
+
  
 
<html><p align="center">
 
<html><p align="center">
<img src="" style="width:400px"  ></a>
+
<img src="https://static.igem.org/mediawiki/2016/4/42/T--SYSU-MEDICINE--BBa_K1993014-fig2.png" style="width:300px"  ></a>
  
 
</p></html>  <br>
 
</p></html>  <br>
  
<p align="center">'''_______________'''</p><br>
+
<p align="center">'''Figure 2 Promoter of α-SMA was activated and green fluorescence could be detected'''</p><br>
  
 
==Reference==
 
==Reference==

Latest revision as of 12:48, 18 October 2016


pSMA-eGFP

eGFP is a type of GFP derivatives by mutation. This mutation dramatically improved the spectral characteristics of GFP, resulting in increased fluorescence, photostability, and a shift of the major excitation peak to 488 nm, with the peak emission kept at 509 nm. (Details could be seen on BBa_K1993017)

Alpha-smooth muscle actin (α-SMA) is the actin isoform that predominates within vascular smooth-muscle cells and plays an important role in fibrogenesis. (Details could be seen on BBa_K1993022)

Previous studies indicated that alpha-smooth muscle actin (α-SMA) could be induced in inflammatory environment and had a relationship with the risk of MSC fibrogenesis, which weakened their therapeutic effect. [1] As a result, we desired to find out a special way to detect the expression of α-SMA in MSCs.

To achieve it, we designed a plasmid with promoter of SMA (pSMA) linking eGFP. (Figure 1)


Figure 1 pSMA-eGFP


To confirm the function of this plasmid, we administered TGF-β1 (7ng/mL). After that, Promoter of α-SMA was activated and green fluorescence could be detected in MSCs (Figure 2)


Figure 2 Promoter of α-SMA was activated and green fluorescence could be detected


Reference

[1] Talele N P, Fradette J, Davies J E, et al. Expression of α-smooth muscle actin determines the fate of mesenchymal stromal cells[J]. Stem cell reports, 2015, 4(6): 1016-1030.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1763
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 887