Zebrafish CRISPR Injection Mix
with sgRNA Validation

All the Power of CRISPR: Complete CRISPR Injection Mix & sgRNA Validation

CRISPR zebrafish injection mix

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, 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 a project 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.

Is CRISPR Injection Mix the right choice for you?

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 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 validated injection mix for researchers looking for a fully plug and play format.

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. 

To mitigate failure rates, our custom injection mix includes an expertly designed set of reagents and tools that can help you:

  • Build tools without the hassle. We put all the moving parts together for you to create the consistency in sourcing and design of the reagents.
  • Improve success rate. Our custom injection mix packages incorporate sgRNA validation/cutting efficiency to mitigate failure rates.
  • Enable do-it-yourself. Our team will do the heavy lifting of the project design and reagent procurement so you can take advantage of CRISPR/Cas9 technology.
  • Save time. You can eliminate lengthy and variable workflow leading up to experiment start time.
  • Jumpstart your project. You select how much help you need without having to be a CRISPR expert.

Injection Mix and sgRNA Validation Offerings

Looking for an easy way to bring CRISPR gene editing into your lab? Choose the level of service that is appropriate for you.

Standard Injection Mix Standard Injection Mix + Screening Primers Validated sgRNA Injection Mix Complete Validated Injection Mix
Design
Injection Mix
Screening Reagents
sgRNA Testing
In vivo Editing Efficiency Screening
Pricing*$995$1,995$3,995$5,995
Timeline*3-4 weeks4-5 weeks4-6 weeks6-8 weeks
More DetailsMore DetailsMore DetailsMore Details
*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.

What do I need?

In order to use our injection mix, you must have access to:

  • Microinjection rig and capabilities for early embryo injection. 
  • Facilities for zebrafish growth, maintenance, and crossing
  • PCR machines and reagents.

Injection Mix and sgRNA Validation Details

Standard Injection Mix

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 by indels. We send a custom designed injection mix and you develop your own preferred detection/screening protocol.

What’s Included?
  • Consultation with a genetic engineer to develop the genome editing strategy
  • Sequence files for the donor homology construct and the final, edited locus
  • In silico design of all reagents including the sgRNA site and donor homology template
  • 4 vials of 10uL lyophilized sgRNAs duplexed with the Cas9 protein
  • Rhodamine dye for early visualization of injection success
  • Detailed Instructions on mix reconstitution and injection protocols.

Standard Injection Mix + Screening Primer

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. 

What’s Included?
  • Our Standard Injection Mix PLUS:
  • Expertly designed PCR primers prescreened for specificity in zebrafish tissue.
  • An in vitro assay to help you optimize detection of your desired edit in your own lab

Validated sgRNA Injection Mix

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?
  • Our Standard Injection Mix PLUS:
  • Selection of two sgRNAs for in vivo evaluation of cutting efficiency
  • Donor homology designed around a demonstrated high efficiency sgRNA site
  • In vivo screening for potential polymorphisms in your target locus

Complete Validated Injection Mix

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 option for complete novices!

What’s Included?
  • Our Validated sgRNA Injection Mix PLUS:
  • We test your injection mix in vivo and determine the somatic integration efficiency to help you decide how many animals to raise.
  • Sequence confirmation of integration capacity of your edit into the zebrafish genome
  • Fully validated PCR primers and protocols for edit detection assay in your own lab.

Injection Mix Service Component Descriptions

Injection Mix

Injection Mix

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 in a stable dehydrated format.

Four injection mixes are provided that reconstitute to 5ul of mix each.

  • Custom reagents from validated suppliers
  • Four 5uL lyophilized injection mixes with instructions on reconstitution
  • Rhodamine dye for early visualization of injection success
Screening Reagents

Screening Reagents

Integration efficiency in injected zebrafish embryosYou 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.

  • PCR primers designed and tested to identify the target edit
  • PCR protocol to identify animals that contain your edit
sgRNA Testing

sgRNA Testing

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.

  • Up to 3 sgRNAs tested for CRISPR cutting efficiency in zebrafish embryos
  • Selection against sgRNAs that have toxic effects
  • Identification of an sgRNA that is successful for guiding CRISPR cutting
In Vivo Editing Efficiency Screening

In Vivo Editing Efficiency Screening

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.

  • Test injection of mix into embryos
  • Integration efficiency in injected embryos
  • Sequencing to confirm precise edit
READY TO GET STARTED?

Ready to connect with us to learn more about working with our company or our technology?

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