Transfection of Capsaspora owczarzaki, a close unicellular relative of animals

Abstract

How animals emerged from their unicellular ancestor remains a major evolutionary question. New genome data from the closest unicellular relatives of animals have provided important insights into the evolution of animal multicellularity. We know that the unicellular ancestor of animals had an unexpectedly complex genetic repertoire, including many genes that are key to animal development and multicellularity. Thus, assessing the function of these genes among unicellular relatives of animals is key to understanding how they were co-opted at the onset of the Metazoa. However, such analyses have been hampered by the lack of genetic tools. Progress has been made in choanoflagellates and teretosporeans, two of the three lineages closely related to animals, whereas no tools are yet available for functional analysis in the third lineage: the filastereans. Importantly, filastereans have a striking repertoire of genes involved in transcriptional regulation and other developmental processes. Here, we describe a reliable transfection method for the filasterean Capsaspora owczarzaki. We also provide a set of constructs for visualising subcellular structures in live cells. These tools convert Capsaspora into a unique experimentally tractable organism to use to investigate the origin and evolution of animal multicellularity.

Patterns of Ancestral Animal Codon Usage Bias Revealed through Holozoan Protists

“…2016 ). Transgenics systems are currently being developed for holozoan protists ( Parra-Acero et al. 2018) , and the data presented here provide important information for the expression of artificial transgenes.…”

Genetic tool development in marine protists: emerging model organisms for experimental cell biology

“…Genomic and transcriptomic data are available for multiple representatives characterized by diverse cell morphologies, some of which can even form multicellular structures 46 . Until recently, only transient transformations had been achieved for some opistokonts such as the filasterean Capsaspora owczarzaki 48 , the ichthyosporean Creolimax fragrantissima 49 and the choanoflagellate Salpingoeca rosetta 21 . Through the EMS initiative, we report on evidence for transient transformation of the ichthyosporean Abeoforma whisleri, isolated from the digestive tract of mussels, and review a recently published stable transformation protocol for S. rosetta achieved by using the selectable puromycin N-acetyl-transferase gene (Fig.…”

Integrin-mediated adhesion in the unicellular holozoanCapsaspora owczarzaki

“…Initial descriptions of Capsaspora showed that it has dynamic filopodia (Stibbs et al, 1979), and a recent study revealed that these filopodia contact the substrate and appear to hold the cell body above the surface (Parra-Acero et al, 2018). We examined filopodia behaviour in live cells, by performing time-lapse microscopy of adherent Capsaspora cells transfected with a membrane marker (NMM-mCherry) (Parra-Acero et al, 2018). We observed that filopodia not only hold the cell over the surface but appear to actively participate in its motility ( Figure 2D).…”