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Protocol for imaging and quantifying single-cell CDK activity levels during early mouse embryonic development by

Bechara Saykali1, Andy D Tran2, Michael J Kruhlak2

  • 1Laboratory of Genome Integrity, CCR, NCI, NIH, Bethesda, MD, USA.

STAR Protocols
|September 9, 2025
PubMed
Summary

This study presents a method to measure cyclin-dependent kinase (CDK) activity in single mouse embryo cells. The protocol enables tracking CDK activity during early development using live imaging and computational analysis.

Keywords:
Bioinformaticscell differentiationdevelopmental biology

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Area of Science:

  • Developmental Biology
  • Cell Biology
  • Molecular Biology

Background:

  • Quantifying protein activity in single cells during early embryonic development is challenging.
  • Cyclin-dependent kinases (CDKs) are crucial regulators of the cell cycle in pre-implantation embryos.

Purpose of the Study:

  • To provide a detailed protocol for measuring cyclin-dependent kinase (CDK) activity in single cells of mouse pre-implantation embryos.
  • To enable researchers to track CDK activity dynamics throughout early embryonic development.

Main Methods:

  • Utilizing fluorescent-based reporters for CDK activity.
  • Employing in vitro culture strategies for live imaging of mouse embryos.
  • Implementing live fluorescent confocal microscopy for image acquisition.
  • Developing computational workflows for cell tracking and data analysis.

Main Results:

  • The protocol allows for the quantification of CDK activity at the single-cell level.
  • Efficient live imaging and cell tracking strategies are described.
  • A comprehensive image analysis workflow for processing and visualization is detailed.

Conclusions:

  • This approach simplifies the complex task of measuring CDK activity in developing mouse embryos.
  • The protocol facilitates the study of cell cycle regulation during pre-implantation development.
  • This method provides valuable insights into the role of CDKs in early embryogenesis.