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Gene knock-in (KI) is the process of insertion of an exogenous sequence into a specific locus of the genome. We use it to introduce stable genetic modifications in the zebrafish genome in order to generate complex disease models and tools boosting their phenotypic characterization.
This technique can be used for introducing in the zebrafish genome short nucleotide sequences (like point mutations, loxP sites or protein tags) or long sequences of exogenous DNA (like reporter genes or mutated genes’ CDSs).
According to your needs, either F1 heterozygous KIs (carrying one exogenous allele) or F2 homozygous KIs (with two exogenous alleles) can be generated and shipped worldwide.
Highly efficient and precise integration of the exogenous sequence of interest into the zebrafish genome.Extremely accurate recapitulation of expression patterns of sequences of interest compared to random transgenesis.
Highly efficient and precise integration of the exogenous sequence of interest into the zebrafish genome.
Extremely accurate recapitulation of expression patterns of sequences of interest compared to random transgenesis.
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.