The Point Mutation service using zebrafish introduces a small number of nucleotide changes at a target site using CRISPR/Cas9. With this service you can:
- study a disease-causing mutation
- humanize a critical amino acid
- explore the binding site of an enzyme
- introduce phosphomimetics
- mutate isoform start sites or make any specific mutation of interest.
We can successfully repair a point mutation by knocking in a wild type repair template to restore gene function. Below is a visual readout of knock-in repair in F0 (injected, generation zero) zebrafish larvae; to determine if this repair has been transmitted through the germline, we would have to raise them to adults and screen their progeny for development of pigment cells.
nacre 48 hpf: a nacre mutant larvae, homozygous loss of function mutation in the gene mitfa (microphthalmia-associated transcription factor a) causing a loss of melanophores (pigment cells)
wt 48 hpf: a wild type larvae with normal development and distribution of melanophores
ODN rescue 48 hpf: nacre mutant embryos injected with Cas9 + gRNA + ssODN (single stranded oligodeoxynucleotides) showing repair of the mitfa mutation, resulting in rescued development of melanophores to varying degrees (highest to lowest degree of rescue, top to bottom)
|Service Package||Price||Est. Delivery Time|
|Full Build||$26,955||9 - 12 months|
|Verified Clutch||$9,310||4 - 6 months|
|Custom Injection Mix (without verification of sgRNA cutting)||$995||2 - 4 weeks|
|Custom Injection Mix (with verification of sgRNA cutting)||$3,259||4 - 8 weeks|