Fig. 1.

Hydractinia Eef1a gene structure and strategy for mutagenesis by homology directed repair. a Genomic organization of Hydractinia Eef1a. Boxes indicate exons, while angled lines indicate introns. Black areas are untranslated regions, white areas are coding sequences. The four exons are indicated as E1, E2, E3, and E4. b Overview of Cas9-mediated insertional mutagenesis. Injection of Cas9:sgRNA complexes and a plasmid containing a DNA repair template leads to the insertion of a FLAG-P2A-eGFP cassette via homology directed repair. c Nucleotide level view of insertional mutagenesis strategy. The single letter codes for the amino acid translation of the coding sequence are shown above the double-stranded DNA sequence. Bracketed text indicates the FLAG-P2A-eGFP cassette. Base pairs shown in italics were inserted to maintain the full Eef1a peptide