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Mosaic Zebrafish Transgenesis for Functional Genomic Analysis of Candidate Cooperative Genes in Tumor Pathogenesis

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BEAM: a combinatorial recombinase toolbox for binary gene expression and mosaic analysis.

Luciano C Greig, Mollie B Woodworth, Alexandros Poulopoulos

Biorxiv : the Preprint Server for Biology

|February 24, 2023

Summary

We developed Binary Expression Aleatory Mosaic (BEAM), a novel method for genetic mosaic analysis in mice. This technique enables comparative studies by creating two distinct cell populations for gene mis-expression or deletion analysis.

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Area of Science:

Developmental Biology
Genetics
Molecular Biology

Background:

Genetic mosaic analysis is crucial for understanding cellular functions but challenging in mice due to complex breeding strategies.
Existing methods for generating genetically distinct cell populations in mice are often time-consuming and labor-intensive.

Approach:

Developed Binary Expression Aleatory Mosaic (BEAM), a system utilizing sparse recombinase activation for generating two intermingled, genetically distinct cell populations.
BEAM relies on combinatorial recombinase activity following DNA construct delivery (transfection or viral transduction) to label cells with distinct fluorescent proteins (e.g., green or red).
Allows for targeted gene mis-expression or deletion in one cell population for direct comparison with adjacent wild-type control cells.

Key Points:

The BEAM system is extensively optimized and validated both in vitro and in vivo.
Demonstrates utility in investigating cell-autonomous functions and identifying temporal or spatial phenotypes.
Effective for analyzing changes in cell proliferation and death, and for controlling experimental variability.

Conclusions:

BEAM offers a powerful and efficient alternative for genetic mosaic analysis in mice.
Facilitates comparative studies of gene function in a mixed-cell environment.
Enhances the investigation of complex biological processes at the cellular level.