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High-throughput, image-based screening of pooled genetic-variant libraries.

ABSTRACT: We report a high-throughput screening method that allows diverse genotypes and corresponding phenotypes to be imaged in individual cells. We achieve genotyping by introducing barcoded genetic variants into cells as pooled libraries and reading the barcodes out using massively multiplexed fluorescence in situ hybridization. To demonstrate the power of image-based pooled screening, we identified brighter and more photostable variants of the fluorescent protein YFAST among 60,000 variants.

SUBMITTER: Emanuel G

PROVIDER: S-EPMC5958624 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature

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High-throughput, image-based screening of pooled genetic-variant libraries.

Emanuel George G Moffitt Jeffrey R JR Zhuang Xiaowei X

Nature methods 20171030 12

We report a high-throughput screening method that allows diverse genotypes and corresponding phenotypes to be imaged in individual cells. We achieve genotyping by introducing barcoded genetic variants into cells as pooled libraries and reading the barcodes out using massively multiplexed fluorescence in situ hybridization. To demonstrate the power of image-based pooled screening, we identified brighter and more photostable variants of the fluorescent protein YFAST among 60,000 variants. ...[more]

PMID: 29083401

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