Part Collection
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.
Figure.1 Schematic diagram of constructs of Left ITR-CMV-AR185-T2A-EGFP- 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-T2A-EGFP-Poly(A) | 1989 | |
| BBa_K2865010 | Composite | CMV-AR185-T2A-EGFP-Poly(A) | 2135 | |
| BBa_K2865012 | Composite | Left ITR-BNP-AR185-T2A-EGFP-Poly(A)-Right ITR | 2387 | |
| BBa_K2865013 | Composite | Left ITR-CMV-AR185-T2A-EGFP-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
- 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.
- 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.
- 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.
- 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.
