The development of CRISPR for a mollusc establishes the formin Lsdia1 as the long-sought gene for snail dextral/sinistral coiling

ABSTRACT The establishment of left-right body asymmetry is a key biological process that is tightly regulated genetically. In the first demonstration in a mollusc of successful germline transmission of a CRISPR/Cas9-edited gene, we show decisively that the actin-related diaphanous gene Lsdia1 is the single maternal gene that determines the shell coiling direction of the freshwater snail Lymnaea stagnalis. Biallelic frameshift mutations of the gene produced sinistrally coiled offspring generation after generation, in the otherwise totally dextral genetic background. This is the gene sought for over a century. We also show that the gene sets the chirality at the one-cell stage, the earliest observed symmetry-breaking event linked directly to body handedness in the animal kingdom. The early intracellular chirality is superseded by the inter-cellular chirality during the 3rd cleavage, leading to asymmetric nodal and Pitx expression, and then to organismal body handedness. Thus, our findings have important implications for chiromorphogenesis in invertebrates as well as vertebrates, including humans, and for the evolution of snail chirality. This article has an associated ‘The people behind the papers’ interview. Highlighted Article: CRISPR/Cas9 reveals that Lsdia1 is the gene that determines the body handedness of Lymnaea stagnalis from the undifferentiated one-cell stage, through the decisive 3rd cleavage and via the nodal and Pitx pathways.