Gene knockouts are a commonly used tool for biologists to understand gene function. Examination of phenotypes when the gene is deleted can reveal insights into what role that gene plays in the organism.
Using CRISPR/Cas9 in zebrafish, we can disrupt integral regulatory domains of a gene or the coding sequence of a gene (KO null), or even remove the entire functional domian or the entire coding sequence depending on gene size (KO deletion).
If deletion of your gene is lethal, consider making a floxed gene.
Standard Injection Mix
Validated sgRNA Injection Mix
Fully Validated Injection Mix
Mosaic Clutch* (F0 injected embryos)
Full Build** (Sequence verified hetero zygous line)
KO Design
Injection Mix Assembly
Locus Evaluation & Sequencing Primers
in vivo sgRNA Testing
in vivo Editing Assessment & Screening Reagents
Expertly Injected Embryos
Germline Transmission Screening & Line Propagation
* This service is only available to clients in the United States.
** Full Build includes screening n=100 adults; standard husbandry charge applies.
# Includes evaluation of 2 sgRNAs
## Includes evaluation of 4 sgRNAs
Choose this option if you want us to pick your sgRNA site, have already tested your sgRNA in vivo, or want to make a CRISPR knockout. We send a custom designed injection mix and you develop your own preferred detection/screening protocol.
What’s Included
Choose this option if you want us to guarantee your injection mix is designed around a high efficiency sgRNA site using our rigorous in-house testing process.
What’s Included?
Choose this option if you want a CRISPR knockin with completely validated reagents to use in your lab and to find your gene edited line. This is the best Custom Injection Mix option for the novice zebrafish line builder.
What’s Included?
Choose this option if you want a CRISPR knockin with completely validated reagents to use in your lab and to find your gene edited line. This is a good option for the researcher who wants to outsource part of the workload in the development of a new line.
What’s Included?
Choose this option if you want us to deliver a stable sequence validated CRISPR knockin line.
What’s Included?
We create a mix using quality custom reagents from our validated suppliers. The sgRNAs are duplexed with the Cas9 protein.
The mix is created using concentrations of Cas9, sgRNA and donor homology that work well in our genome editing pipeline. The mix is provided as a stable dehydrated reagent.
Four injection mixes are provided that reconstitute to 5ul of mix each.
You can miss your edit if you don’t have a good assay to detect it.
We will design and test a robust assay to detect your edit, which is critical when you begin germline screening.
We will also provide PCR primers and protocols for an edit detection assay.
Ensure that you have the quality reagents you need for a successful genome edit. The efficiency of the sgRNA guided cutting can vary widely and is directly tied to the achievement of a successful genome edit.
Algorithms and experience help us choose the best candidate sgRNAs for your project, but the guided cutting efficiency of an sgRNA can be impacted by genome organization, for example, chromatin accessibility. The best way to determine the effectiveness of an sgRNA in a Zebrafish genome is to test it in vivo in the injected embryos.
We test your sgRNAs for their ability to guide Cas9 cutting. Our process also identifies SNPs and polymorphoisms in your locus that may impact homology directed repair. Embryos are injected with two different sgRNA/Cas9 duplexes and we determine the cutting efficiency for each sgRNA. If both sgRNAs fail to guide successful cutting, a third sgRNA is injected and tested.
Additional sgRNA testing can also be added with consultation.
Have total peace of mind by having us test your injection mix in our facility.
Our skilled injectionists will inject embryos with your mix and determine the somatic integration efficiency by PCR.
We will confirm the precise edit by sequencing to ensure that your edit is occurring correctly with no erroneous insertions.
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Built on a robust portfolio of CRISPR genome editing patents, InVivo Biosystems has established a comprehensive preclinical discovery and development platform. By leveraging alternative models and proprietary methods, we accelerate go-to-market timelines for pharmaceutical companies and beyond.
