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Mosaic analysis with double markers in mice.

Hui Zong1, J Sebastian Espinosa, Helen Hong Su

  • 1Department of Biological Sciences, Stanford University, Stanford, CA 94305, USA.

Cell
|May 11, 2005
PubMed
Summary

Mosaic analysis with double markers (MADM) in mice enables simultaneous cell labeling and gene knockout. This technique efficiently labels cells in vivo and reveals developmental fates, such as cerebellar granule cell progenitor lineage determination.

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Area of Science:

  • Developmental biology
  • Genetics
  • Neuroscience

Background:

  • Studying cell lineage and gene function in vivo requires precise labeling and manipulation techniques.
  • Existing methods often lack the ability to simultaneously label and knockout genes in specific cell populations.

Purpose of the Study:

  • To introduce a novel method, Mosaic Analysis with Double Markers (MADM), for simultaneous labeling and gene knockout in vivo.
  • To demonstrate the efficiency and broad applicability of MADM in various cell types and tissues.
  • To utilize MADM to investigate cell fate determination in the developing cerebellum.

Main Methods:

  • MADM involves knocking in two reciprocal chimeric genes at identical chromosomal locations.
  • Functional marker expression and recombination are induced by Cre recombinase.
  • The method allows for the creation of labeled clones of somatic cells for lineage tracing and gene knockout studies.

Main Results:

  • MADM efficiently induces interchromosomal recombination in both mitotic and postmitotic cells across all examined tissues.
  • The study successfully created conditional knockouts in small, labeled cell populations.
  • MADM revealed that cerebellar granule cell progenitors are specified early, with their progeny exhibiting restricted axonal projection patterns within the cerebellar cortex.

Conclusions:

  • MADM is a powerful and versatile tool for in vivo cell labeling, gene knockout, lineage tracing, and neuronal connection studies.
  • The method provides new insights into developmental processes, exemplified by the early fate determination of cerebellar granule cell progenitors.
  • MADM significantly advances the ability to study gene function and cell behavior at the clonal level in complex biological systems.