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Updated: Apr 11, 2026
Genetic Engineering of Dictyostelium discoideum Cells Based on Selection and Growth on Bacteria
Published on: January 25, 2019
A large-scale screen reveals genes that mediate electrotaxis in Dictyostelium discoideum
Runchi Gao1, Siwei Zhao2, Xupin Jiang3
1School of Life Sciences, Yunnan Normal University, Kunming, Yunnan 650500, China. Departments of Dermatology and Ophthalmology, Institute for Regenerative Cures, School of Medicine, University of California at Davis, Davis, CA 95817, USA. Department of Cell Biology and Anatomy, Johns Hopkins University, School of Medicine, Baltimore, MD 21205, USA.
Abstract:
Directional cell migration in an electric field, a phenomenon called galvanotaxis or electrotaxis, occurs in many types of cells, and may play an important role in wound healing and development. Small extracellular electric fields can guide the migration of amoeboid cells, and we established a large-scale screening approach to search for mutants with electrotaxis phenotypes from a collection of 563 Dictyostelium discoideum strains with morphological defects. We identified 28 strains that were defective in electrotaxis and 10 strains with a slightly higher directional response. Using plasmid rescue followed by gene disruption, we identified some of the mutated genes, including some previously implicated in chemotaxis. Among these, we studied PiaA, which encodes a critical component of TORC2, a kinase protein complex that transduces changes in motility by activating the kinase PKB (also known as Akt). Furthermore, we found that electrotaxis was decreased in mutants lacking gefA, rasC, rip3, lst8, or pkbR1, genes that encode other components of the TORC2-PKB pathway. Thus, we have developed a high-throughput screening technique that will be a useful tool to elucidate the molecular mechanisms of electrotaxis.
