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Fig. S6 Assessing CRISPR-Cas9-mediated knockout in zebrafish injected with RetroMyelin gRNA, related to Figure 7 and STAR Methods (A and B) Example chromatograms from Sanger sequencing of a PCR product amplified from genomic DNA extracted from a control-injected larva (A) or RetroMyelin gRNA-injected larva (B) Predicted genomic sequence indicated on top, and gRNA target and PAM sequence indicated by red box and arrow. Below the traces are predicted alleles identified from the chromatograms using CRISP-ID,64 showing only few SNPs in control but multiple indels and variations in the gRNA-injected larva. (C and D) Alignments of the sequencing results for PCR products amplified from genomic DNA extracted from several control (C) or RetroMyelin gRNA-injected (D) larvae to the predicted genomic sequence (top, gRNA sequence in blue, and predicted cut site in red). Note widespread mismatches in RetroMyelin gRNA-injected traces, indicating gRNA activity.
Reprinted from Cell, 187, Ghosh, T., Almeida, R.G., Zhao, C., Mannioui, A., Martin, E., Fleet, A., Chen, C.Z., Assinck, P., Ellams, S., Gonzalez, G.A., Graham, S.C., Rowitch, D.H., Stott, K., Adams, I., Zalc, B., Goldman, N., Lyons, D.A., Franklin, R.J.M., A retroviral link to vertebrate myelination through retrotransposon-RNA-mediated control of myelin gene expression, 814830.e23814-830.e23, Copyright (2024) with permission from Elsevier. Full text @ Cell