From 2007.igem.org

How It works

1) Myosin Heavy Chain (MHC) is exposed on infarcted heart tissue.

2) scFv chimeric receptor targeting MHC, binds to a repeated domain of MHC.

The scFv chimeric receptor contains a fusion of the following protein domains: anti-MHC-Fc, EporD2, EporTM, and gp130i, dnaEC-VP16. EpoR= erythopoeitin receptor. dnaEC= carboxy domain of the Sp Synechosystis split intein.

3) EporD2 facilitates the dimerization of the chimeric receptors

4) A mutant version of EporTM inhibits ligand independent dimerization of the chimeric receptors

5) gp130i becomes activated and JAK's phosphorylate tyrosines on gp130i, including Y759.

6) Shp2-mLexA-dnEN binds to phosphorylated Y759 of gp130i. (mLexA is a mutated form of E coli DNA binding protein LexA that does not contain a cryptic nuclear localization sequence found in the wild-type).

7) Shp2-mLexA-dnEN comes in close proximity to the dnaEC-VP16 fused to the c-terminus of the gp130i

8) dnaEN and dnaEC, parts of the Sp Synechosystis split intein system, undergo peptide processing resulting in the release of newly constituted fusion transcription factor mLexA-VP16.

9) mLexA-VP16 translocates to the nucleus where it binds to minimal promoters containing lexA operators to switch on expression of cell-cell interaction proteins such as N-Cadherin , anti-apoptotic proteins such as Bcl-XL,p35 and IAP and blood vessel growth factors such as VEGF.