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Conditional Genetic Transsynaptic Tracing in the Embryonic Mouse Brain

Conditional Genetic Transsynaptic Tracing in the Embryonic Mouse Brain

Published on: December 22, 2014

Evaluation of TRAP-sequencing technology with a versatile conditional mouse model.

Mike Hupe¹, Minerva Xueting Li, Karin Gertow Gillner

¹Ludwig Institute for Cancer Research Ltd, Box 240, Stockholm SE-171 77, Sweden, Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm SE-171 77, Sweden and Max-Planck-Institute for Molecular Biomedicine, Department of Tissue Morphogenesis, and University of Münster, Faculty of Medicine, D-48149 Muenster, Germany.

Nucleic Acids Research

|October 30, 2013

Summary

Researchers developed a new mouse model, mCherryTRAP, for in vivo translational profiling of specific cell types. This tool aids in understanding gene expression in rare cells, like endothelial cells, and improves data analysis for cell differentiation studies.

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Area of Science:

Molecular Biology
Genetics
Developmental Biology

Background:

In vivo molecular profiling of rare cell populations, such as endothelial cells, presents significant challenges.
Existing methods for translating ribosome affinity purification (TRAP) require robust filtering strategies to isolate cell type-specific transcripts.
Differential RNA expression between the translatome and transcriptome can complicate data interpretation.

Purpose of the Study:

To develop a versatile Cre-dependent translating ribosome affinity purification (TRAP) mouse model for lineage-specific translational profiling.
To address the challenges of in vivo molecular profiling for rare and interspersed cell populations.
To evaluate the implications of translatome-transcriptome differences for TRAP studies and demonstrate applicability to endothelial cell differentiation.

Main Methods:

Generation of a versatile floxed translating ribosome affinity purification (TRAP) mouse model, termed mCherryTRAP.
Application of the mCherryTRAP model for Cre-dependent translational profiling of distinct cell lineages from intact tissues.
Analysis of differential RNA expression between the translatome and transcriptome in embryonic brains and kidneys.

Main Results:

The mCherryTRAP mouse model enables Cre-dependent translational profiling of specific cell lineages from intact tissues.
Extensive differential expression of RNAs between the translatome and transcriptome was observed in embryonic brains and kidneys.
The study demonstrated the utility of the mCherryTRAP model for studying organ-specific endothelial cell differentiation.

Conclusions:

The mCherryTRAP mouse model provides a powerful tool for in vivo translational profiling of diverse cell populations, including rare types.
Understanding translatome-transcriptome differences is crucial for accurate data interpretation in TRAP studies.
This technology facilitates the investigation of cell differentiation processes at the translational level.