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As an officially CRISPR licensed provider, ZeClinics can generate tailored knock-in (KI) lines by introducing an exogenous sequence into a specific locus of the zebrafish genome (site-directed mutagenesis).
Using the CRISPR/Cas9 system in zebrafish, we can insert short nucleotide sequences (like point mutations, loxP sites, or protein tags) or long sequences of exogenous DNA (like reporter genes or mutated genes’ CDSs).
This strategy is often used to understand gene function, to more precisely model human diseases, or to establish genetic tools to support phenotypic analysis.
Get the edit you want along with the support you need to start your KI project.
Compared to Tol2 transgenesis, CRISPR/Cas9 is less efficient but much more precise. Unlike, overexpressing genes by mRNA injection, KI induces stable and permanent edits in the zebrafish genome.
Targeted integration of the exogenous sequence of interest into the zebrafish genomeExtremely accurate recapitulation of the expression patterns of the target genesProduce disruption in WT sequence
Targeted integration of the exogenous sequence of interest into the zebrafish genome
Extremely accurate recapitulation of the expression patterns of the target genes
Produce disruption in WT sequence
To obtain the targeted insertion of an exogenous DNA sequence, one cell stage zebrafish embryos are injected with Cas9/sgRNA complexes targeting the gene of interest and a donor DNA containing the sequence of the KI allele. Successful integration can be proved either by deep sequencing, KI-specific PCR, or by expression of fluorescent reporters. Injected fish are grown to sexual maturity and subsequently screened to identify a founder carrying the KI allele in its germline.
If you want more information about our knock-ingeneration services or have any other questions,please contact our experts.