XB-ART-53822Sci Rep. November 15, 2017; 7 (1): 4858.
The Cannabinoid Receptor Interacting Proteins 1 of zebrafish are not required for morphological development, viability or fertility.
PubMed ID: 28687732
PMC ID: PMC5501828
Article link: Sci Rep.
Genes referenced: cnr1 cnrip1
Article Images: [+] show captions
|Figure 1. Cnrip1 genes in zebrafish. (a) Amino acid sequence alignment of Cnrip1a and Cnrip1b with the two splice variants of the single human CNRIP1 gene, named CRIP1a and CRIP1b. Distinct exons are shown in alternating black and blue font (red indicates amino acids coded by two exons). Comparing both zebrafish genes and CRIP1a asterisk indicates identity and colon similarity between all three, full stop similarity between any two. Bold indicates identity between CRIP1a and CRIP1b. (b) Synteny of adjacent genes in vertebrate CNRIP1 loci. Colours link homologous genes.|
|Figure 3. Mutagenesis of cnrip1a and cnrip1b. (a) Alignments of wild type with each of three mutant alleles showing the predicted expressed mutant polypeptide beneath. Bold underline indicates the gRNA target, red font the protospacer motif recognised by Cas9, hyphens deleted bases, blue highlight inserted bases and asterisks novel stop codons. In the mutant protein sequences, bold text indicates the residual wild type fragment and normal font the aberrant polypeptide tail. (b,c). To test for nonsense-mediated decay, about 50 siblings from in-crosses of cnrip1a +/kg98 (b) and cnrip1b +/kg101 (c) mutant carriers were subjected to in situ hybridisation for the cognate mRNA at the indicated stages, photographed, DNA extracted, PCR performed across the mutant locus and genotype confirmed by DNA sequencing as indicated beneath each panel. Quantitative evidence of reproducibility is given in Table S1.|