Zebrafish Genome-Editing Services

Design Construct Deliver Custom Gene-Edited Zebrafish

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We are a zebrafish CRO that offers multiple gene-editing services including but not limited to mutagenesis, tagging, and injection.

Our scientific team has 40 years of combined zebrafish gene editing experience and access to the start-of-art University of Oregon Zebrafish Facility and top-rated zebrafish researchers. We use CRISPR and Tol2 as our genome editing methods.

Our service can significantly reduce your experiment design time and can be tailored to different experimental requirements.

Introducing Our New Services

  • Point mutation knock-in using CRISPR/Cas9: Create genetic models using zebrafish for understanding gene function, basic biology, and more precisely modeling human diseases. Learn more >>
  • Complete CRISPR Injection Mix: Looking for an easy way to bring CRISPR gene editing into your lab? Choose the level of service that suits you or your lab the best, ranging from standard ready-to-inject mixes to full evaluation of sgRNA cutting efficiency. Learn more >>

Site-Directed Mutagenesis

Using CRISPR/Cas9 technology, we can modify zebrafish genome through Knockout, Point Mutation, Precise Deletion, Floxed Allele, etc. Specific experiment requirements can be met by working with our expert design team members. Learn More »

Knockout

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, we can delete integral domains or the entire the coding sequence of a gene in zebrafish, depending on gene size.

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Point Mutation

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.

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Precise Deletion

The Precise Deletion service removes a small defined region of DNA via CRISPR/Cas9 gene editing in zebrafish. This can be useful for deleting protein domains, DNA regulatory regions, or any sequence of your choice.

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Floxed Allele

CRISPR/Cas9 is used to insert loxP sites flanking the region to be deleted in zebrafish. This line can then be crossed with a Cre-line expressing the Cre recombinase under a specific promoter or injected with a plasmid containing Cre recombinase. The Cre recombinase promotes recombination of the two loxP sites and the region between the sites is removed from the genome.

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Endogenous Tagging

We can help you visualize protein expression and help you conduct biochemical studies of your gene of interest using CRISPR/Cas9. Custom tags and combinations(i.e. GFP::3xFLAG, etc) are also available. Learn More »

Fluorescent Tag

The addition of a fluorescent protein tag at the endogenous locus enables you to visualize your protein in vivo without altering the level of gene expression. In Zebrafish, fluorescent proteins are added using CRISPR/Cas9 gene editing technology. You can choose from a variety of protein tags including YFP, BFP, GFP, and mCherry.

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Epitope Tag

The addition of an epitope tag at the endogenous locus enables you to quantify your protein without altering the level of gene expression. Epitope tags in zebrafish are added using CRISPR/Cas9 gene editing technology. You can choose from a variety of tags including FLAG, HA, HIS, TAP, or S-peptide.

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Expression

Obtain gene knock-ins (KIs) in zebrafish with expression at random insertion loci via Tol2. Learn More »

Tol2

Insert large content in the zebrafish genome using the Tol2 transposon-based system. Using Tol2 transgenesis you can generate: Cre Recombinase-expressing lines; Transcription reporters; Models to analyze tissue-specific gene expression; Models to control the timing of gene expression; Humanized zebrafish. Insertion content can range between 2 – 10 kb.

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Complete CRISPR Injection Mix

Reduce time from experiment conception to execution with the affordable Complete CRISPR Injection Mix package.

All The Power of CRISPR & More

Our Complete CRISPR Injection Mix includes Cas9 protein, an RNA-based sgRNA(s), and a donor homology construct and can be shipped as a lyophilized mix in as little as 2 weeks. Each Complete CRISPR Injection Mix project includes: consultation; sequence files for the donor homology construct and the final, edited locus; development of all reagents including the sgRNA and donor homology construct; 2 – 10 uL lyophilized injection mixes with instructions on reconstitution

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Custom Design Project

Cannot find what you are looking for? Contact us at support@invivobiosystems.com

Zebrafish Genome Editing Service Packages

We offer a variety of service packages to fit the needs and budget of your lab. Choices range from the fast and affordable Complete CRISPR Injection Mix to the comprehensive Full Build service package, allowing you to customize the components you need to accomplish your gene editing goals.

Price reflects academic pricing

Service
Cost
Delivery Time
Content
Benefits
INJECTION MIX
$995 and up
Min. 3 weeks
Reagents
Make genome edits in yours lab-specific strains.
VERIFIED CLUTCH*
$9,310 and up
Min. 4 months
Injected clutch of embryos
Expert design and pre-screening on a budget.
FULL BUILD
$26,995 and up
Min. 9 months
Germline-verified fish
Receive animals ready to cross with your strains.
*Verified Clutch service is only available to customers in the United States.

Key References

A well rounded review provides a detailed roadmap outlining -numerous tools available for genetic modification in zebrafish, including the historical and experimental context in which they have been used. Methods including Tol2 transgenesis, TALENs and CRISPR/Cas9 are discussed. A particularly useful flowchart can help readers understand the numerous decision points and considerations involved in choosing the right approach for their particular experimental needs.

Download the publication.

An excellent framework for the novice and expert genome engineer alike.The article outlines the design of a number of elegant genetic tools to facilitate CRISPR/Cas9 mediated knock-ins, as well as downstream screening and experimental uses. The focus here is on fluorescent tags, epitope tags, conditional tools, and transgenesis markers to help researchers screen for phenotypically silent edits. 

Download the publication.

If you’re looking for more mechanistic insight into gene editing in zebrafish, look no further than this review from Simone et al. The authors provide a detailed discussion of the different DNA repair pathways, template types, and experimental considerations that all factor into a successful gene editing endeavor. The various “designer nucleases” available for zebrafish gene editing applications, their advantages, and their limitations are also discussed.

Download the publication.

Process For Generating Zebrafish Lines

InVivo Biosystems provides a streamlined zebrafish gene editing service. Getting customized zebrafish lines made easy.

  • Zebrafish Project Design

    Project Design

  • sgRNA-test-zf

    sgRNA Testing

  • Zebrafish Donor Homology

    Donor Homology Design

  • Zebrafish Injection

    Injection

  • Zebrafish Clutch Verification

    Clutch Verification

  • Zebrafish Verification

    Germline Verification

  1. Project Design:

    Selection of best guide RNAs surrounding the locus to be edited.
  2. sgRNA Testing:

    In vivo cutting assayed by PCR/sequencing to determine the most efficient sgRNA.
  3. Donor Homology Design:

    Donor homology plasmid or DNA oligonucleotide is designed based on the best sgRNA.
  4. Injection:

    Injection of mix into zebrafish embryos at the 1 cell stage.
  5. Clutch Verification:

    Verification of edit by PCR and/or sequencing of 10% of injected embryos.
  6. Germline Verification:

    Verification of germline editing.

Features of Our Services

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    CRISPR

    Gene Editing

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    Zebrafish

    Advantageous Model Organism

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    Service Packages

    Choice of Simply or Complex Build

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    Expert Service Team

    With Over 40 Years of Experience

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    Timely Delivery

    With Ensured Quality

Why Outsource to InVivo Biosystems For Zebrafish Gene-Editing Service

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Reduction in Experimental Design Time

NemaMetrix helps you save design time and facilitates your progress.

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Custom Modification for Specific Needs

Work with us to design a workflow tailored for your requirements.

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Long Years of Passion and Innovation

40 years of experience makes us the best in this field.

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Quality Standard

Our specialist are highly trained to gurantee the products and solutions are well certified

Frequently Asked Questions

First, you need to inform us your gene of interest for gene editing so we can provide you with information regarding what we will be able to assist. Second, you need to choose one service package of interest. It can be full build, verified clutch, or customer injection mix. Depending on your choice, time of preparation varies.

Complete Injection Mix is simple to work with, all you need to do is add water, inject, and screen. Our Complete CRISPR Injection Mix includes Cas9 protein, an RNA-based sgRNA(s), and a donor homology construct and can be shipped as a lyophilized mix in as little as 2 weeks. Discuss your research goals with us, and NemaMetrix will provide you with an expertly designed injection mix to generate your new zebrafish line.

Depending on the option you made, we either have a 3 step service payment plan or a full payment requirement. If you choose our Full build or Verified Clutch Projects. The 3 step payment is applied, The Phase I cost is required for project start and is non-refundable. Upon completion of Phase I, the Phase II cost is due and is also non-refundable. Phase III is due at the end of the project and is only billable if a germline-integrated line is delivered. Prices stated are exclusive of all taxes, fees, licenses, duties or levies and, unless otherwise stated in the quotation, transportation charges, freight and insurance. If you choose our Complete CRISPR Injection Mix Project, full payment is required before the start of the project.

Using Tol2 transgenesis you can generate Cre Recombinase-expressing lines, transcription reporters, models to analyze tissue-specific gene expression, models to control the timing of gene expression, and humanized zebrafish. Insertion content can range between 2 – 10 kb.

Depending on each individual country’s import regulations & guidelines. We can ship embryos, larvae, juvenile and adult fish as long as they are allowed in the receiving country.

Case Studies

See how NemaMetrix is helping scientists with zebrafish gene editing.

Mutations in EYS are known to cause photoreceptor degeneration in patients with RP25, but functions of the EYS protein in the vertebrate retina are poorly understood.

ABC research lab aims to study the function of Eye shut homolog for survival of photoreceptors in zebrafish. They proposed several method of mutagenesis in EYS and found out that CRISPR/Cas9 is the best technique. However, due to lack of experience in practicing CRISPR/Cas9, ABC researchers had difficult time introducing mutations of EYS gene on the cone photoreceptors. After several unsuccessful trials, ABC researchers have still not found out a functional protocol to knockout EYS gene. While searching for the protocols for zebrafish CRO to outsource, ABC researchers discovered the zebrafish gene-editing service offered by NemaMetrix.

NemaMetrix zebrafish service team planned out a detailed workflow for zebrafish EYS gene knockout, and carried out the entire project in a turn-key fashion. After 6 months, the genetically modified zebrafish was delivered to the lab, after which observations were made every week on the development of retinal photoreceptors. A deficiency of EYS protein was observed in all 200 individuals, suggesting a 100% knockout success rate.

Progressive degeneration of cones and rods were discovered to correlate with decreased level of EYS protein. ABC derived a solid conclusion from this research and published paper after confirming their prediction with the help of NemaMetrix’s zebrafish gene editing service.

B pharmaceutical company designed a drug de novo aiming to treat Reticular Dysgenesis and advanced to in vivo testing of the de novo drug in model organisms.

After intense debate on which model organisms should be chosen, zebrafish was determined to be used considering its relative advantage to model human diseases especially pediatric genetic diseases. However, B company is not experienced in genetically modifying zebrafish genome, so the research team decides to seek for assistance from a zebrafish CRO that specialize in gene editing. They browsed several zebrafish CROs and finally decided on outsource NemaMetrix due to its experience and credential.

After discussion about this case, both companies decided on using GFP tagging to indicate the expression level of AK2 gene before and after usage of de novo drug, and different dosages were applied to discover the optimum daily usage. Level of expression were determined by the measuring the amount of fluorescence.

The drug was discovered to be effective in raising the expression of AK2 proteins, indicted by the increasing level of GFP detected after drug injection. B company was able to proceed to the next stage of testing drug side effect.

A zebrafish research lab in University of C hoped to figure out the differential expression of a oncogenic gene KRAS in different types of tissue.

A main focus of this experiment is to see how the nervous system can be affected by KRAS mutation. Researchers have already decided that Cre-lox system would be a best fit in this scenario.

Due to limited knowledge of zebrafish gene editing, the researchers sought assistance from other zebrafish research labs, during which they obtained information about several zebrafish CROs, and finally agreed to outsource NemaMetrix. They were interested by the TOL2 transposon-based system as well as the efficiency service delivery.

Out of three options, verified was chosen due to short preparation period. The modified zebrafish were delivered three months after the order, and subsequent experiment was successful. Using Cre-Recombinase inducing drugs, tissue specific loss of function mutation of KRAS was achieved by the researchers.

Observations were made and they drafted a paper describing their findings.

D lab was working on discovering cancer promoting genes. They managed to target a human gene named THOR which facilitate the growth of melanoma in human. In order to validate their hypothesis, researchers in this lab picked zebrafish as an experiment model.

Zebrafish is known to be an advantageous model to study human disease. The team concentrated on discovering human oncologic genes, and was not able to spend too much time on the validation process, especially on reagent design and validation. However, as a crucial part of the experiment, D lab cannot derive a conclusion without testing their result.

D lab did a research online and found out that NemaMetrix had a suitable option for them to outsource which is the complete CRISPR injection mix.

With this option, they only need to provide NemaMetrix with the THOR gene construct, and the injection mix could be shipped back to them in as short as 2 weeks, including Cas9 protein, an RNA-based sgRNA(s), and a donor homology construct as a lyophilized mix. Lab D started the process right way and received the injection mix in two weeks. They then injected the mix into 100 different zebrafish, which are chemically induced to develop melanoma.

They found out that THOR incorporation into the zebrafish genome induced faster tumor growth in the model, and thus proved their hypothesis to be correct.

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