Composite

Part:BBa_K4963001

Designed by: Aniruddha Atul Nayak, Sumiran Kasturi   Group: iGEM23_BITSPilani-Goa-India   (2023-10-12)


Cannabinoid Receptor 1 (Modified) + INPN

The composite part is a surface expressing drug receptor for cannabinoid targets, comprising a truncated Cannabinoid Receptor 1 and Ice nucleation protein derivative INP - N. The modified CB1 domain functions as a drug receptor. The short INP - N domain functions as a surface expression vehicle which is transported to the outer regions.
The composite part serves the purpose of a surface-based holding device on the cell for a drug (particularly honokiol, the primary drug in our project), thereby administering a recombinant cell vector as the carrier which further functions as an effective locator of the tumor site and further localizes in the region. Therefore, the part allows for an effective drug delivery and tumor dispersion system, for the purpose of our project. The part in general may be used as a device for drug delivery via cellular vectors.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1077
    Illegal PstI site found at 1776
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1077
    Illegal PstI site found at 1776
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 333
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1077
    Illegal PstI site found at 1776
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1077
    Illegal PstI site found at 1776
    Illegal NgoMIV site found at 75
    Illegal NgoMIV site found at 408
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1575


At the surface, the N - terminal of the domain binds to the surface lipid glycosylphosphatidylinositol (GPI), thus forming a secure anchor.1 The CB1 domain is carried to the surface simultaneously.
The CB1 domain comprises the prominent binding sites for Honokiol. This domain functions as a holding and releasing agent for the drug. Release activity is observed in the presence of AM - 6538 (the primary antagonist in our project). The reduced size of the part further enhances competitive interaction between the primary drug and the antagonist.



Fig. 1 The composite part design and functioning. Created with BioRender.


Source Parts
1. CB-1 (modified): https://parts.igem.org/Part:BBa_K4963000
2. INP - N: https://parts.igem.org/Part:BBa_K3279006


References
1. Li L, Kang DG, Cha HJ. Functional display of foreign protein on surface of Escherichia coli using N-terminal domain of ice nucleation protein. Biotechnol Bioeng. 2004 Jan 20;85(2):214-21. doi: 10.1002/bit.10892. PMID: 14705004.


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Categories
Parameters
None