Extended Data Fig. 3: CRISPR-Cas9 knockout of Ciona Hmx and Ciona Ngn.

a. Placement of sgRNA guides for Hmx CRISPR knockout and primers used for validation, relative to gene structure. Guide and primer sequences in Methods and Supplementary Table 1. b. Predicted engineered outcome of Hmx CRISPR knockout. c. PCR amplification of Ciona intestinalis (Type B) genomic DNA from wild type, CRISPR control and Hmx CRISPR embryo DNA (as well as from additional sgRNAs that were tested but not used in further experiments). The guide used in further experiments is marked in yellow. Sizes of bands in the DNA ladder (100bp DNA-Ladder, extended: Carl Roth) are given in base pairs (bp). d. Sequencing of amplified bands with sequence identity matching the predicted outcomes in (b). e. Placement of sgRNA guides for Ngn CRISPR knockout and primers used for validation, relative to gene structure. Guide and primer sequences in Methods and Supplementary Table 1. f. Predicted engineered outcome of Ngn CRISPR knockout. g. PCR amplification of Ciona intestinalis (Type B) genomic DNA from CRISPR control and Ngn CRISPR embryo DNA (as well as from additional sgRNAs that were tested but not used in further experiments). The guide used in further experiments is marked in yellow. Sizes of bands in the DNA ladder (100 bp DNA-Ladder, extended: Carl Roth) are given in base pairs (bp). h. Sequencing of amplified bands with sequence identity matching the predicted outcomes in (f).