We can create a floxed allele for tissue- or temporal- specific deletion of your gene of interest. This can be very useful when studying embryonic lethal genes or for understanding how your gene of interest functions in different tissues.
CRISPR/Cas9 is used to insert loxP sites flanking the region to be deleted. This line can then be crossed with a Cre-line expressing the Cre recombinase under a specific promoter or injected with a plasmid containing Cre recombinase. The Cre recombinase promotes recombination of the two loxP sites and the region between the sites is removed from the genome.
One of the more difficult aspects of biology is how to perform loss-of-function studies of essential genes. Now the conditional mutagenesis by loxP insertion in zebrafish is possible. At NemaMetrix, we have turned to sequential integration of loxP sequences in two stages of injection and screening to circumvent both large deletions across loxP target loci as well as the homology-independent integration of repair templates at opposing target sites. Learn more.
- Used to make deletion of genes in site and time specific manner
- Flank gene of interest with LoxP sites
- Inject a plasmid or cross into strain that expresses Cre recombinase where/when/how you want loss-of-function to occur
|Service Package||Price||Est. Delivery Time|
|Full Build||$27,365||18 months|
|Verified Clutch||Not Available|
|Custom Injection Mix (without verification of sgRNA cutting)||$1,990||2 - 4 weeks|
|Custom Injection Mix (with verification of sgRNA cutting)||$4,254||4 - 8 weeks|