Part:BBa_K2520014

CMV-Mono display:MOG 3-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) is a tag that binds 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 multiple sclerosis (BBa_K2520038) and is component related to our specific project. The CMV promoter (BBa_K1119006) is a constitutive expression promoter in mammalian cells and hGH (BBa_K404108) serves as a terminator.

MS disease

In MS, the immune system attacks the protective sheath (myelin) that covers nerve fibers and causes communication problems between your brain and the rest of your body. Eventually, the disease can cause the nerves themselves to deteriorate or become permanently damaged.

In MS disease, T cells attack 3 types of glycoprotein:

1) Myelin basic protein (MBP)

2) Proteolipid protein (PLP)

3) myelin oligodendrocyte glycoprotein (MOG)

Experimental autoimmune encephalomyelitis (EAE):

EAE is a CD4' T cell-mediated inflammatory disease of the central nervous system (CNS) induced in experimental animals with CNS homogenate, myelin or myelin components. In its chronic form, EAE is a well accepted experimental model for multiple sclerosis (MS).

Epitopes:

MOG glycoprotein is the only CNS autoantigen known to induce both a T-cell response and a demyelinating autoantibody response in EAE

Therefore, we chose the three epitopes that induced the most significant immune response. MOG3 is the third epitope that appears in the MOG glycoprotein sequence (304-337)

References

Mendel, Itzhack, Nicole Kerlero de Rosbo, and Avraham Ben‐Nun. "A myelin oligodendrocyte glycoprotein peptide induces typical chronic experimental autoimmune encephalomyelitis in H‐2b mice: Fine specificity and T cell receptor Vβ expression of encephalitogenic T cells." European journal of immunology 25.7 (1995): 1951-1959.‏ 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 614
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1313