Coding

Part:BBa_K3113010

Designed by: Theresa Keil   Group: iGEM19_Munich   (2019-09-04)
Revision as of 23:15, 21 October 2019 by Theresakeil (Talk | contribs)


MCP

MS2 Coat Protein (MCP) is a capsid protein that self-assembles to a capsid. The protein acts as an adapter for RNA binding into extracellular vesicles. The protein binds to the MS2-stemloop and loads mRNA into the extracellular vesicles.

Usage

The informational readout of ALiVE is RNA. To load a specific RNA into the vesicles we fused it to RNA-motifs which are bound specifically by RNA binding proteins. Two combinations of binding protein and motif were tested. One was MCP, the MS2 bacteriophage coat protein, which binds to the MS2 RNA motif.

Biology

The coat protein of the RNA bacteriophage MS2 binds a specific stem-loop structure in viral RNA to accomplish encapsidation of the genome and translational repression of replicase synthesis. [1] Bacteriophage MS2 is an icosahedral virus with 180 copies of a coat protein forming a shell around a single-stranded RNA molecule.[2]


Characterization

Western Blot

Figure 1:

Vesicle Export in HEK293T

Figure 2: Extended version with more CD63 constructs shown.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

  1. ↑ Peabody, D S. “The RNA binding site of bacteriophage MS2 coat protein.” The EMBO journal vol. 12,2 (1993): 595-600.
  2. ↑ The refined structure of bacteriophage MS2 at 2.8-A resolution. Golmohammadi R., Valegaard K., Fridborg K., Liljas L. J. Mol. Biol. 234:620-639(1993)
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