Tech Spec. Team Superfly!
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1 Tech Spec Team Superfly!
2 Remyelination
3 Impact Numerous diseases deal with demyelination, and symptoms are severe! Greatly improve quality of life for many individuals 2.1 million people worldwide with Multiple Sclerosis (200 new cases every week in US alone) Method of treatment applicable to many diseases
4 Our Solution Engineer a bacterial cell that will: o Recognize demyelinated axons once in the brain o Adhere to these axons o Secrete myelin o Unadhere from the axon once it is appropriately myelinated Input: Bare axon; Output: Myelin
5 Device Level Description 1. Axon-cell adhesion device 2.Axon recognizing device 3.Myelinating device
6 Device Diagram
7 No Myelin?
8 Already Myelinated?
9 Timing Diagram
10 Chassis Clostridium Sporogenes o o o o Motile, anaerobic bacteria Part of the C.Botulinum family, but do not produce neurotoxins Currently being used in cancer research to reduce tumor growth The spores of the bacteria are small enough to reach the brain and can grow in anaerobic conditions
11 Axon-Cell Adhesion Device Cell-Axon Adhesion Device CADM4 CADM3
12 Axon-Cell Adhesion Device CADM4-CADM3 adhesion pair (CADM = Cell Adhesion Molecule) In PNS: CADM4 = Schwann cells, CADM3 = unmyelinated axon Gene: CADM4 o Promoter : HG_ACW:41758 o Transcript: CADM4.DAPR07-UNSPLICED Debugging: IGSF4C antibodies (N-term)
13 Axon Recognizing Device Axon Recognizing Device F3/contactin CADM4 L1 protein CADM3
14 Axon Recognizing Device F3/Contactin cellular receptor L1 (axon) binds to F3/contactin (oligodendrocytes) F3/contactin and b1-integrin stimulate activation of Fyn Fyn normally phosphorylates protein hnrnp A2 and begins a signalling cascade Gene: CNTN1 F3/Contactin: Immunoglobulin glycoprotein B1-integrin: Transmembrane receptors that provide information from the extracellular matrix Fyn: A tyrosine kinase molecule
15 Control Mechanism Function: Turn the production of myelin ON when the chassis recognizes the demyelinated axon The fyn protein can bind to the HIV LTR promoter to induce the transcription of myelin genes Debugging: place HIV LTR promoter on GFP rather than myelin genes, place in isolated cell, record if cell turns green
16 Myelin Producing Device F3/contactin Myelin Producing Genes CADM4 L1 protein CADM3 Myelin Proteins
17 Myelin Producing Device Three main glycoproteins produced by Schwann cells: 1. Myelin Protein Zero (Po Protein) 2. Myelin-Associated Glycoprotein (MAG) 3. Peripheral Myelin Protein-22 (PMP-22)
18 Myelin Protein Zero Encoded by the Myelin Protein Zero gene (MPZ) The most abundant protein in myelin Acts like an adhesion and protective molecule Location: o Schwann cells o Chromosome 1 (Position 23.3)
19 Myelin-Associated Glycoprotein Encoded by the MAG gene Axon-Glia interactions Maintenance of myelin with aging Location: o Myelinating Schwann Cells
20 Peripheral Myelin Protein-22 Encoded by peripheral myelin protein-22 gene (PMP-22) Plays a crucial role in the development and maintenance of myelin Location: o Primarily in Schwann Cells o Chromosome 17 (Position 12)
21 Myelin Producing Device Debugging: Isolate myelin genes with promoter and insert into cell Provide the cell with FYN in culture Observe whether or not myelin is produced
22 Things to look into Effects o immune system o off-target o self-destruction? Structure o Superfly vs. natural myelin o myelinating genes o myelin leaving the cell Debugging o wrapping around axon o feedback
23 Go/No Go
24 References E47F635775E64919DA02016C185E0.y7ESLndDIsN8cE7qwvy6w%3a
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