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The injection of zebrafish one-cell-stage embryos with Cas9/sgRNAs complexes makes it possible to induce genomic mutations at the desired site with very high efficiency (up to 100%).
Thanks to this extremely high performance, at ZeClinics we are able to generate somatic mutants (Crispants) carrying a high rate of biallelic mutations in F0 animals. This approach ensures the creation of “de facto” F0 mutants enabling an exhaustive evaluation of loss-of-function phenotypes in a short time (2-3 months). Genotyping of mosaic F0 larvae permits a fine genotype/phenotype correlation, enabling the achievement of extremely reliable results. In addition, a Crispants-based approach is the ideal system to evaluate, in vivo, the activity of new tools (like modified or improved Cas9 proteins).
High throughput screening of multiple candidate genesFast target validation prior to the generation of stable KONon-invasive, in vivo readouts to detect KO-induced phenotypes at the level of cells, tissues or organs Unlike larvae injected with morpholino or antisense oligonucleotides, Crispants do not recover a normal gene expression after the first days of developmentPrecise genotype/phenotype correlation
High throughput screening of multiple candidate genes
Fast target validation prior to the generation of stable KO
Non-invasive, in vivo readouts to detect KO-induced phenotypes at the level of cells, tissues or organs
Unlike larvae injected with morpholino or antisense oligonucleotides, Crispants do not recover a normal gene expression after the first days of development
Precise genotype/phenotype correlation
We are an officially licensed CRISPR solutions provider. We design sgRNAs having high specificity and efficiency to guarantee a mutagenesis rate close to 100%. A specific sgRNA guides the Cas9 endonuclease to the desired locus of interest to induce targeted double-strand breaks. Error-prone mechanisms of DNA-repair can cause the generation of small insertions and deletions (INDELs) potentially disrupting the coding sequence of the targeted gene and promoting the formation of loss-of-function mutations.
To generate mutant alleles in zebrafish, the Cas9 and the sgRNA(s) designed against the selected gene are injected into wild-type one-cell-stage embryos (F0). Mutagenesis takes place during early stages of embryonic development, giving rise to F0 larvae carrying a mosaic set of somatic and germline mutations. High mutagenesis rates in somatic KOs enable the evaluation of the resulting loss-of-function phenotypes already in F0 larvae.
Phenotypes resulting from the genetic modification can be characterized on demand in the whole organism or in specific organs.
Results of the phenotypic analysis according to customers' interests and needs: