Team:NMU China/Parts

Combatting COVID-19
Naval Medical University CHINA

Basic Parts

We designed 17 basic parts in total.You can click on the part number to see the details of basic parts.

NameType Description
BBa_K4040000 Protein_Domain Intracellular Domain of the MEGF10 Protein
BBa_K4040001 Protein_Domain Intracellular Domain of the MERTK Protein
BBa_K4040002 Protein_Domain FcRγ
BBa_K4040003 Protein_Domain CD3 zeta chain
BBa_K4040004 Protein_Domain CD8 hinge
BBa_K4040005 Tag Myc tag
BBa_K4040006 Protein_Domain TEV protease
BBa_K4040007 Protein_Domain CD8TM
BBa_K4040011 Protein_Domain tTA
BBa_K4040013 Protein_Domain mIL-6R
BBa_K4040014 Protein_Domain CD8a signal peptide
BBa_K4040019 Protein_Domain CR3022 scFv
BBa_K4040032 Protein_Domain IL-6 signal transducer, gp130
BBa_K4040034 Protein_Domain Truncated HER2
BBa_K4040035 Protein_Domain Truncated EGFR
BBa_K4040036 Protein_Domain CD20
BBa_K4040037 Protein_Domain T2A peptide
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Composite Parts

We designed 10 composite parts in total to constitute our project. You can click on the part number to see the details of composite parts.

NameType Description
BBa_K4040015 Composite CAR-MEGF10
BBa_K4040016 Composite CARγ
BBa_K4040017 Composite CAR ζ
BBa_K4040018 Composite CAR-MERTK
BBa_K4040020 Composite GP130-TEV
BBa_K4040021 Composite IL6R-TCS-Gal4-KRAB
BBa_K4040022 Composite IL6R-TCS-tTA
BBa_K4040023 Composite pSV40-UAS-GFP Expression Cassette
BBa_K4040024 Composite pTET-mCherry-PGK-CD20 Expression Cassette
BBa_K4040033 Composite UAS-pSV40

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Favourite Parts

Our favorite Basic Part is CD3 zeta chain (BBa_K4040003), which encodes for the CAR-CD3Z intracellular segment. This part successfully transmits CAR-perceived signals into the cell, triggering subsequent cascades that activate CAR-M phagocytosis and induce the release of inflammatory factors. CD3Z intracellular segment has been widely used in CAR-T intracellular segment, and intracellular segment for CAR-M has also been reported in previous literature. All CD3 chains contain immunoreceptor tyrosine-based activation motifs (ITAMs) in their cytoplasmic domain. Upon TCR engagement, these motifs become phosphorylated by Src family protein tyrosine kinases LCK and FYN, resulting in the activation of downstream signaling pathways [1,2].
Future iGEM teams working with CAR can make use of it to create CAR-Ts and CAR-Ms which induce phagocytosis and immune response. As seen in our results, CARzeta macrophages phagocytosed Spike-bearing 293 cells in an S-specific manner. CARzeta mediated significantly stronger levels of SARS-CoV-2 phagocytosis by THP-1 cells than CARD(truncated CAR).
Our favourite composite part is CARMERTK (BBa_K4040018), which was regarded as an unsuccessful receptor in a previous report [3] and showed no cellular killing effect on target cells when expressed in THP-1 cells in our assay, demonstrated a virion clearance capacity similar to that of CARγand CARzeta. Our data further support that CARMERTK mediates ‘immunologically silent' virion removal, which does not elicit a proinflammatory response. It opens the possibility for future iGEM teams to build such CAR without strong inflammatory response.

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Improvement of existing parts

We made some improvements based on BBa_K3132016 from SMMU_China-2019 to reduce the background expression activity of the receptor. You can click on the part number to see the details of improvement.

NameType Description
BBa_K4040025 Protein_Domain GEMS
BBa_K4040026 Protein_Domain GEMS with one Alanine added
BBa_K4040027 Protein_Domain GEMS with two Alanines added
BBa_K4040028 Protein_Domain GEMS with three Alanines added
BBa_K4040029 Protein_Domain GEMS with four Alanines added

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[1]Barber EK, Dasgupta JD, Schlossman SF, Trevillyan JM, Rudd CE. The CD4 and CD8 antigens are coupled to a protein-tyrosine kinase (p56lck) that phosphorylates the CD3 complex. Proc Natl Acad Sci U S A. 1989 May;86(9):3277-81. doi: 10.1073/pnas.86.9.3277. PMID: 2470098; PMCID: PMC287114.
[2]Iwashima M, Irving BA, van Oers NS, Chan AC, Weiss A. Sequential interactions of the TCR with two distinct cytoplasmic tyrosine kinases. Science. 1994 Feb 25;263(5150):1136-9. doi: 10.1126/science.7509083. PMID: 7509083.

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