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Phenomics-Based Quantification of CRISPR-Induced Mosaicism in Zebrafish
Claire J Watson1, Adrian T Monstad-Rios1, Rehaan M Bhimani1
1Department of Orthopaedics and Sports Medicine, University of Washington, Seattle, WA, USA; Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA.
Cell Systems
|March 20, 2020
Summary
CRISPR screens in zebrafish reveal how somatic mutations cause mosaicism, mimicking human genetic diseases like osteogenesis imperfecta. This study offers a new method to analyze these spatially variable phenotypes.
Area of Science:
- Genetics
- Developmental Biology
- Zebrafish Models
Background:
- Genetic mosaicism leads to varied phenotypes within an organism.
- CRISPR-Cas9 gene editing allows for precise genetic modifications.
Purpose of the Study:
- To quantitate spatially variable phenotypes in CRISPR-edited zebrafish.
- To understand the biological basis of mosaicism from somatic mutations.
Main Methods:
- Imaging-based phenomics to analyze phenotypes in the zebrafish axial skeleton.
- Characterization of loss-of-function cell clusters and their size distribution.
- Quantification of phenotypic mosaicism in somatic mutants of plod2 and bmp1a genes.
Main Results:
- Identified a distinctive size distribution of cell clusters arising from clonal fragmentation and merger.
- Demonstrated phenotypic convergence between somatic G0 mutants and homozygous germline mutants.
- Developed statistical frameworks for analyzing spatial phenotypic variation.
Conclusions:
- CRISPR screens in G0 zebrafish can accurately model human genetic diseases.
- This study provides a novel approach to decode spatially variable phenotypes in mosaic organisms.
