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)

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

Figure2

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