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Versatile phenotype-activated cell sorting.
Lee, Jihwan; Liu, Zhuohe; Suzuki, Peter H; Ahrens, John F; Lai, Shujuan; Lu, Xiaoyu; Guan, Sihui; St-Pierre, François.
Afiliação
  • Lee J; Systems, Synthetic, and Physical Biology Program, Rice University, Houston, TX 77005, USA.
  • Liu Z; Department of Electrical and Computer Engineering, Rice University, Houston, TX 77005, USA.
  • Suzuki PH; Department of Bioengineering, Rice University, Houston, TX 77005, USA.
  • Ahrens JF; Department of Bioengineering, Rice University, Houston, TX 77005, USA.
  • Lai S; Department of Neuroscience, Baylor College of Medicine, Houston, TX 77030, USA.
  • Lu X; Systems, Synthetic, and Physical Biology Program, Rice University, Houston, TX 77005, USA.
  • Guan S; Department of Neuroscience, Baylor College of Medicine, Houston, TX 77030, USA.
  • St-Pierre F; Systems, Synthetic, and Physical Biology Program, Rice University, Houston, TX 77005, USA. stpierre@bcm.edu.
Sci Adv ; 6(43)2020 10.
Article em En | MEDLINE | ID: mdl-33097540
ABSTRACT
Unraveling the genetic and epigenetic determinants of phenotypes is critical for understanding and re-engineering biology and would benefit from improved methods to separate cells based on phenotypes. Here, we report SPOTlight, a versatile high-throughput technique to isolate individual yeast or human cells with unique spatiotemporal profiles from heterogeneous populations. SPOTlight relies on imaging visual phenotypes by microscopy, precise optical tagging of single target cells, and retrieval of tagged cells by fluorescence-activated cell sorting. To illustrate SPOTlight's ability to screen cells based on temporal properties, we chose to develop a photostable yellow fluorescent protein for extended imaging experiments. We screened 3 million cells expressing mutagenesis libraries and identified a bright new variant, mGold, that is the most photostable yellow fluorescent protein reported to date. We anticipate that the versatility of SPOTlight will facilitate its deployment to decipher the rules of life, understand diseases, and engineer new molecules and cells.

Texto completo: 1 Base de dados: MEDLINE Idioma: En Ano de publicação: 2020 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Idioma: En Ano de publicação: 2020 Tipo de documento: Article