Part:BBa_K1993014
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1763
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 887
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