Establishment of Transgenesis in the Demosponge Suberites domuncula

Genetics
Roger Revilla-I-DomingoFlorian Raible

Abstract

Sponges (Porifera) represent one of the most basally branching animal clades with key relevance for evolutionary studies, stem cell biology, and development. Despite a long history of sponges as experimental model systems, however, functional molecular studies are still very difficult to perform in these animals. Here, we report the establishment of transgenic technology as a basic and versatile experimental tool for sponge research. We demonstrate that slice explants of the demosponge Suberites domuncula regenerate functional sponge tissue and can be cultured for extended periods of time, providing easy experimental access under controlled conditions. We further show that an engineered expression construct driving the enhanced green fluorescence protein (egfp) gene under control of the Suberites domuncula β-actin locus can be transfected into such tissue cultures, and that faithfully spliced transcripts are produced from such transfected DNA. Finally, by combining fluorescence-activated cell sorting (FACS) with quantitative PCR, we validate that transfected cells can be specifically reisolated from tissue based on their fluorescence. Although the number of detected enhanced green fluorescent protein (EGFP)-expressing cells is ...Continue Reading

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Citations

Jul 29, 2019·Wiley Interdisciplinary Reviews. Developmental Biology·Jeffrey Colgren, Scott A Nichols
Jun 5, 2019·Nature Protocols·Alexander KlimovichThomas C G Bosch
Sep 8, 2021·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Bernd SchierwaterRob DeSalle

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Methods Mentioned

BETA
transfection
PCR
Fluorescence-Activated Cell Sorter
electrophoresis
FACS
Fluorescence
fluorescence microscopy
electron microscopy
transgenic
single-cell sequencing

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