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iGEM Berkeley iGEMBerkeley iGEMBerkeley

Many applications in synthetic biology demand precise control over subcellular localization, cell morphology, motility, and other such phenotypes that are only observable via microscopy. At present, engineering these properties is challenging due in large part to the inherent throughput limitation imposed by microscopy. We have developed a strategy that enables high-throughput library screening with microscopy by coupling a unique fluorescence signature with each genotype present in a library. These MiCodes (microscopy barcodes) are generated by targeting combinations of fluorophores to several organelles within yeast, and they eliminate the need to isolate and observe clonal populations separately. MiCodes can potentially scale to library sizes of 10^6 or more, and their analysis can be largely automated using existing image processing software. As a proof of principle, we applied MiCodes to the problem of finding unique pairs of protein-protein interaction parts.

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Localization tags, promoter optimization, and promoter characterization.

MiCode construction using Golden Gate Assembly.

Automation techniques for image acquisition and cell identification with Matlab and Cell Profiler.

Leucine Zippers affinity assay.

The UC Berkeley iGEM team would like to thank Agilent and the Synthetic Biology Institute for their financial support, Synberc for their administrative support, and IDT for discounted oligos.