RT Journal Article SR Electronic T1 Genome editing enables reverse genetics of multicellular development in the choanoflagellate Salpingoeca rosetta JF bioRxiv FD Cold Spring Harbor Laboratory SP 2020.02.18.948406 DO 10.1101/2020.02.18.948406 A1 David S. Booth A1 Nicole King YR 2020 UL http://biorxiv.org/content/early/2020/02/19/2020.02.18.948406.abstract AB In a previous study, we established a forward genetic screen to identify genes required for multicellular development in the choanoflagellate, Salpingoeca rosetta (Levin et al., 2014). Yet, the paucity of reverse genetic tools for choanoflagellates has hampered direct tests of gene function and impeded the establishment of choanoflagellates as a model for reconstructing the origin of their closest living relatives, the animals. Here we establish CRISPR/Cas9-mediated genome editing in S. rosetta by engineering a selectable marker to enrich for edited cells. We then use genome editing to disrupt the coding sequence of a S. rosetta C-type lectin gene, rosetteless, and thereby demonstrate its necessity for multicellular rosette development. This work advances S. rosetta as a model system in which to investigate how genes identified from genetic screens and genomic surveys function in choanoflagellates and evolved as critical regulators of animal biology.