Device

Part:BBa_K2520012

Designed by: Noa Eden, Dana Kadosh   Group: iGEM17_TECHNION-ISRAEL   (2017-10-25)


EF1a-Mono display:pro insulin-hGH

This device is a complete system for displaying proteins on cells' membrane. The IgK leader (BBa_K2520024) is a short signal peptide that prompts the translocation of a protein to the cellular membrane. PDGFR (BBa_K2520026) is a trans-membrane domain that anchors all the components located between the IgK leader and the PDGFR itself to the membrane. HA (BBa_K2520030) and Myc (BBa_K2520031) are tags that bind to specific antibodies and provide an indirect method for verifying the display of proteins on the membrane. The protein that we chose to express on the membrane is an epitope that is known target of diabetes (BBa_K2520027) and is component related to our specific project. The EF1a promoter (BBa_K2520023) is a constitutive promoter in mammalian cells and hGH (BBa_K404108) serves as a terminator. EF1a promoter is very useful in cells where other promoters, such as CMV, are underactive or silenced (such as stem cells).

Diabetes type 1

Diabetes type 1 is a chronic disease of the pancreas that is caused due to T cells attacking the insulin producing beta cells, within the pancreas. Globally, nearly 40 million people suffer from type I diabetes. This disease leads pancreatic dysfunction and insulin dependence.

Proinsulin: Proinsulin is the prohormone precursor to insulin made in the beta cells of the islets of Langerhans. Proinsulin formed by three distinct chains: The A chain, B chain, and the area connecting the two chains - C peptide.

Proinsulin.jpeg

References

(1) Wong, F. Susan, et al. "Identification of an MHC class I-restricted autoantigen in type 1 diabetes by screening an organ-specific cDNA library." Nature medicine 5.9 (1999): 1026-1031.‏

(2) Narendran, Parth, Stuart I. Mannering, and Leonard C. Harrison. "Proinsulin—a pathogenic autoantigen in type 1 diabetes." Autoimmunity reviews 2.4 (2003): 204-210.‏ APA


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 643
    Illegal XhoI site found at 1042
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 777
    Illegal AgeI site found at 151
    Illegal AgeI site found at 1315
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2762


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Categories
Parameters
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