Raw Sanger sequences of the tyr fragment from crispant and control zebrafish embryos

Thanks to the amenability to genetic modifications, the minimal or
non-invasive phenotyping possibilities in early embryos and larvae and the
presence of a high percentage of genes orthologous to humans, zebrafish is
a successful alternative vertebrate to validate new disease genes and
variants and to study mechanisms underlying human diseases. We developed
an optimized method allowing early and gentle genotype detection to reduce
further possible suffering associated with the generation of genetically
modified zebrafish and valid to strategize the employment of “surplus”
animals (e.g., those needed solely for line generation or without the
desired genotype). The method is based on minimally-invasive tissue (fin)
scratching (FS) to obtain genomic DNA material and perform genotyping in
early zebrafish embryos. We showcase the method's usefulness for
various genotyping needs, including screening both F0 and stable
CRISPR/Cas9 lines and obtaining genomic material compatible with Sanger
sequencing. Here we provide the raw sequences obtained from FS-derived
genomic DNA of single mutant F0 embryos carrying mutations in the tyr gene
generated with Base editor (BE) CRISPR/Cas9 technology. We compared the
sequencing outcome of FS- vs whole embryos (WE)-based genomic DNA
preparation (the WE sequencing results are also provided here). Sequences
show the successful C>T (G>A) conversion generating crispant
tyr mutants that FS-derived genomic DNA can capture. The resulting dataset
demonstrates the compatibility of the method developed with
sequencing-based genotyping of early crispant embryos.