Parts

Overview

Our project, Cytopia, aims to construct Saccharomyces cerevisiae strains capable of efficiently expressing the antimicrobial peptide LL-37. During the development process, we designed and utilized a large number of parts, including 50 basic and 40 composite parts, totaling 90 parts.

All parts have been submitted to the iGEM Registry and functionally validated to ensure their reliability for use by the iGEM community. For more detailed information about our parts, we strongly recommend you visit the specific part pages. You can also click here to learn more about how we designed and optimized these parts.

Our favorite composite parts are BBa_251COFY9 and BBa_25NP2VZ8, as they most significantly enhance the production of LL-37 in S. cerevisiae.

Basic Parts

Our basic parts include multiple functional modules, such as hybrid promoter with higher expression efficiency (BBa_25OJHDQH), signal peptides that assist LL-37 secretion (e.g. BBa_K3789005), and modification tags (e.g. BBa_250A92KZ).Based on our CytoFlow model, we also obtained an LL-37 variant (BBa_25UB42UM) with stronger antibacterial activity.

These basic parts provide reliable support for the construction of composite parts.

Composite Parts

Our composite parts were mainly designed for the subcellular localization, secretion, and efficient expression of LL-37. We assembled fluorescent co-localization parts to visualize LL-37 localization in different organelles (e.g. BBa_25NO7917), and characterized the expression efficiency of hybrid promoters using fluorescent proteins (e.g. BBa_25CSLYKP).We also constructed a series of gene knockout parts to reduce production costs by deleting key regulators in the galactose-dependent system (BBa_25KID167), and to lower intracellular degradation of LL-37 by knocking out specific proteases (e.g. BBa_253NFL7O), which further increased LL-37 yield.