Emanuel George, Moffitt Jeffrey R, Zhuang Xiaowei
Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.
Graduate Program in Biophysics, Harvard University, Cambridge, Massachusetts, USA.
Nat Methods. 2017 Dec;14(12):1159-1162. doi: 10.1038/nmeth.4495. Epub 2017 Oct 30.
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.
我们报告了一种高通量筛选方法，该方法能够对单个细胞中的多种基因型和相应表型进行成像。我们通过将带有条形码的遗传变异作为混合文库引入细胞，并使用大规模多重荧光原位杂交读出条形码来实现基因分型。为了证明基于图像的混合筛选的威力，我们在60000个变异中鉴定出了荧光蛋白YFAST更亮且光稳定性更高的变异体。
