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 will need 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|