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Mutant resources for functional genomics in Dictyostelium discoideum using REMI-seq technology
Nicole Gruenheit1, Amy Baldwin2, Balint Stewart1
1Centre for Life's Origins and Evolution, Department of Genetics, Evolution and Environment, University College London, Darwin Building, Gower Street, London, WC1E 6BT, UK.
BMC Biology
|August 25, 2021
Summary
Researchers developed REMI-seq, a method for high-throughput gene function analysis in the social amoeba Dictyostelium discoideum. This technique enables rapid identification of gene functions by linking genetic mutations to observable traits.
Area of Science:
- Cell Biology
- Developmental Biology
- Evolutionary Biology
Background:
- Ascribing gene function is a major challenge despite advances in genome sequencing.
- Genetically tractable model systems are essential for functional genomics research.
- The social amoeba Dictyostelium discoideum is a widely used eukaryotic model organism.
Purpose of the Study:
- To develop high-throughput methods for gene function analysis in Dictyostelium discoideum.
- To create resources for efficient linking of genotype to phenotype.
- To facilitate large-scale parallel phenotypic analyses.
Main Methods:
- Adaptation of Tn-seq called REMI-seq (restriction enzyme-mediated integration sequencing).
- Quantitative identification of drug resistance marker insertion sites.
- Development of a near genome-wide mutant resource and a pool of barcoded mutants.
Main Results:
- REMI-seq enables high-throughput, quantitative identification of insertion sites.
- Creation of a near genome-wide collection of individual mutants.
- Generation of a defined pool of barcoded mutants for parallel analysis.
- Rapid identification of novel regulators of cell migration, phagocytosis, and macropinocytosis.
Conclusions:
- REMI-seq provides powerful tools for high-throughput gene function analysis.
- The developed methods and resources significantly enhance functional genomics in Dictyostelium discoideum.
- Freely accessible resources and data analysis pipelines are available online.
