Parts
Parts
Triple-Negative Breast Cancer (TNBC) represents a significant clinical challenge due to its aggressive nature and lack of targeted therapies. Current diagnostic methods rely on invasive biopsies, while treatments often struggle against the tumor's complex defenses. Our project introduces a novel, two-pronged synthetic biology approach for the non-invasive diagnosis and subsequent immuno-sensitization of TNBC. The first stage of our system employs a modular "tumor tag" engineered to specifically bind to the HN-1 protein, a biomarker overexpressed in the TNBC tumor microenvironment (TME). Fused with a Green Fluorescent Protein (GFP) reporter, this tag enables in-situ detection of cancerous tissue, offering a potential alternative to surgical biopsies. The second, therapeutic stage is designed to dismantle the TME's defenses by simultaneously inhibiting three key pathways that enable tumor growth and immune evasion. By deploying agents targeting VEGF, CTLA-4, and PD-1, our system aims to weaken the tumor, making it vulnerable to the body's natural T-cell response. This strategy seeks to re-empower the patient's own immune system to recognize and combat cancer cells, thereby significantly enhancing the safety and efficacy of existing therapeutic modalities.
Parts: Constitutive Promoter J23100 Ribosome Binding Site B0034 Double Terminator B0015 LldR(TFA) K1847001 CRP, cAMP receptor protein K861160 PLldR M36021 TRans. Fac. pH(TFphoB) phoB from E.coli Constitutive Promoter J23109 Tumor Tag New Part: HN-1 RED Floracence Protein E1010/Part:BBa_K082010 Pasr K1231000 Ribosome Binding Site B0034 α-VEGF K1694003 TFfhlA K778001 PfdhF K387003 Collagen TAG New Part: ModSILY A-DDRs & Self Cleaving peptide New Part Trans. Fac. Hypoxia Complex(HIF-1α + Flexable Linker +HIF-1β) K2295002 + K5415000 Flexible Linker K105012 Hypoxia Response Element(HRE) K1456004 HRE (Hypoxia Response Element) pTal (Minimal Promoter) K2295003 α-PD-1 K2865001 Outer membrane tag K1694002 α-CTLA-4 New Part PcstA (glucose-repressible promoter) K118011 AvLox l-Lactate oxidase New Part SigH New Part P-atp2 (alkali promoter) New Part : P-atp2 K3796204 hSDC4-T2A-STEAP3 K2796025 Promoter PdhR from e.coli NEW PART TF Ppdhr NEW PART GFP E0040
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Figure 1. Incorporation of a custom crRNA spacer into the SynLOCK Cassette.
Figure 2. Upon replacing the reporter with the custom crRNA spacer, colonies lose their purple color, making the screening process fast and straightforward.
