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CRISPR/Cas9 gene editing is a rapidly growing molecular biology toolset with diverse applications. However, designing and executing a CRISPR project turns out to be more complex than it appears from the outset. Reagent sourcing, and design and validation can be a time intensive process requiring strong internal expertise without a guarantee of success.
We know you have tight deadlines, and may have limited hands or expertise. Our goal is to help you get the tools you need to move forward with your experiments and accelerate the time from your idea to execution of the project. You can work with our team of genome editing specialists to design your injection mix so all you have to do is add water, inject and start screening!
Zebrafish Injection Mix: Comparing Buy vs. DIY.
Our CRISPR injection mixes are fully customizable to get you the edit you want along with the support you need to start your project. Our injection mix packages range from standard ready-to-inject mixes to full in vivo evaluation of sgRNA cutting efficiency and in house validated screening tools to accommodate YOUR level of expertise. Our Standard Mix is for experienced researchers with extensive injection and screening competencies; the Complete Validated Mix is a rigorously tested injection mix for researchers looking for a fully plug and play format.
To mitigate failure rates, our custom injection mix includes an expertly designed set of reagents and tools that can help you:
Images below show two different zebrafish lines using the CRISPR Injection Mix. (Left): SelN-BFPSTOP with BFP followed by a STOP codon inserted into the N-terminus of Selenon (SelN). (Center and Right): Ryr1a-mCherry line where the endogenous Ryr1a was tagged with mCherry at the N-terminus. Image courtesy of Melissa A. Wright, MD/PhD, Assistant Professor of Pediatric Neurology at University of Colorado.
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In order to use our injection mix, you must have access to:
Looking for an easy way to bring CRISPR gene editing into your lab? Choose the level of service that is appropriate for you.
*Prices reflect academic pricing.
*Pricing is for point mutation and knockout projects. Projects that require plasmid builds adds $2,000 and 4-6 weeks to each project.
*In-depth consultation with our expert CRISPR designers and editors can be included with any project.
In order to use our injection mix, you must have access to:
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.
Choose this option if you want to inject and screen for your edit by PCR without having to worry about designing your own screening primers. This option includes our standard mix plus ready-to-go screening primers.
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.
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.
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.
Ready to connect with us to learn more about working with our company or our technology?
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An expert in CRISPR genome editing, InVivo Biosystems creates custom genome-edited C. elegans and zebrafish models to enable aging, developmental, and disease studies. Our unique in vivo platforms and technologies bridge the gap between cells and mice.
