Por Mauro Rebelo

Technology proof of concept

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The first contract aimed to deliver a proof of concept (TRL3), which we stipulated as the genetic modification of mussel cells. It was an intense stage of research and, according to ANEEL’s scale of development, classified as ‘applied research’. This is because there are many knowledge gaps in the biochemistry, molecular physiology and cell biology of the golden mussel.

The work began with the search for candidate genes to target for genetic modification. They should be genes that prevent the mussel from reproducing. But information on reproduction genes in bivalves was so scarce that we had to look for analogues (or, more correctly, orthologues) in other species. In the end, we came up with a list of 26 target genes. But this list is changing as our understanding of mussel reproduction increases.

We also needed to test whether existing gene editing tools would work on mussels. A laborious stage of trial and error from which there is no escape. The biggest challenge turned out to be obtaining the germ cells to carry out the modification experiments. However, we have managed to genetically modify spermatozoa to express the green fluorescent protein (GFP) gene and contribute to the understanding that there is transcription in this cell type (a controversial issue in the literature).

At this stage, we also analyzed mussels from 5 reservoirs, from the south to the northeast of the country, using the ddRAD technique, with more than 8000 loci monitored, to conclude that there are no different populations in Brazil and, therefore, all mussels could be impacted by the genetic modification that is being developed.

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Tags: TRL#3