cas9 expression  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Validated CRISPR/Cas9 guide RNAs targeting neurodevelopmental genes in the tunicate Ciona robusta

doi: 10.64898/2026.03.25.711585

Figure Lengend Snippet: A) Selected Tyrosinase sgRNA target sites are indicated, along with Amplicon-EZ (Genewiz) indel plots and estimated mutagenesis rates. Mutagenesis rates are approximated for plots in which naturally occurring indels confounded automatic rate estimation. B) Left: diagram of simple pigmentation assay (see text for details). Right: results of pigmentation assay of Tyr CRISPR larvae using sgRNAs 1.185 and 5.16 together (40 µg each) in combination with 50 µg Sox1/2/3>Cas9::Geminin Nterminus . ( ; ). CRISPR larvae (n = 45) were compared to negative control larvae (n = 29) electroporated with 80 µg of the “control” sgRNA plasmid instead . Plasmid DNA amounts indicated per 700 µl of total electroporation volume. Statistical significance determined using Fisher’s exact test. C) Selected VAChT sgRNA target sites indicated, along with indel plots. Dashed red line indicates alternative splicing of first exon with remaining exons encoding choline acetyltransferase (ChAT) out of view, as part of a conserved cholinergic locus. Red arrows indicate CRISPR-generated indel peaks. Grey asterisks indicate naturally-occurring indels.

Article Snippet: We are also happy to share access to our onboarded sgRNA and Cas9 expression at Twist Bioscience, for easy custom synthesis and cloning of new sgRNAs and Cas9 vectors.

Techniques: Amplification, Mutagenesis, CRISPR, Negative Control, Control, Plasmid Preparation, Electroporation, Alternative Splicing, Generated