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Project Abstract

MiCodes - enabling library screens with microscopy by connecting genotypes to observable phenotypes

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.

The Micodes Library

Intro here... link to more specifications here

Application: Leucine Zippers

Intro here... link to more specifications here

Thanks for reading, etc... for a summary of what we've accomplished, check out our results page.

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 == '''Project Abstract''' ==
-MiCodes (microCodes) are a system of cell barcodes that allow one to determine a yeast cell's genotype from its individual fluorescent phenotype. By localizing fluorescent proteins to distinct organelles within the cell, each yeast variant displays a unique pattern of colors.  As the number of colors and target organelles raises, an exponential increase is found in the possible number of distinct barcodes.  An extensive number of cells can be visualized at once by exciting the fluorescent proteins using fluorescence microscopy. Through computer or visual selection, one can sift through each combinatorial pattern of colors and targeted locations to identify a desired phenotype and elucidate the associated genotype.
+MiCodes - enabling library screens with microscopy by connecting genotypes to observable phenotypes
+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.
 == The Micodes Library ==