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HaloTag-based reporters for sparse labeling and cell tracking.

Lydie Couturier1, Juan Luna1,2, Khalil Mazouni1

  • 14D Unit, Developmental and Stem Cell Biology Dept, Institut Pasteur, CNRS UMR3738, 75015 Paris, France.

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|November 3, 2022
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Summary

Researchers developed a genetic tool for sparse cell labeling in Drosophila tissues. This method enables real-time tracking of individual cell behavior, advancing studies in developmental biology and morphogenesis.

Keywords:
Cell trackingDrosophilaHaloTagcell fatecell shapeimaginal disclive imagingmorphogenesissingle cell analysissparse labelling

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Area of Science:

  • Developmental Biology
  • Cell Biology
  • Genetics

Background:

  • Multiscale analysis of morphogenesis requires real-time tracking of numerous individual cells over extended periods.
  • Automated, large-scale cell tracking in developing embryos and tissues presents significant challenges.

Purpose of the Study:

  • To develop a novel genetic tool for random and sparse labeling of individual cells in developing Drosophila tissues.
  • To overcome limitations of genetically encoded fluorescent proteins in rapidly dividing tissues.

Main Methods:

  • Utilized a genetic tool for conditional expression of a nuclear HaloTag protein.
  • Employed a cell-permeable synthetic ligand for irreversible fluorescent labeling of HaloTag.
  • Developed an HaloTag version of LifeAct to study cell shape changes.

Main Results:

  • Achieved rapid labeling of individual cells in cultured Drosophila imaginal discs using nuclear HaloTag.
  • Demonstrated the utility of sparse labeling for facilitating cell tracking.
  • Successfully produced an HaloTag-LifeAct fusion protein for cell shape analysis.

Conclusions:

  • Nuclear HaloTag reporters offer a valuable method for sparse, rapid cell labeling in Drosophila.
  • This tool facilitates single-cell analysis of fate dynamics in ex vivo cultured Drosophila tissues.
  • The developed system advances the study of morphogenesis and cellular behavior in developing organisms.