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CRISPR/Cas9 Zebrafish Knockout Services For Precise Genome Editing

CRISPR/Cas9 gene editing has revolutionized the field of genome engineering. InVivo specializes in providing highly accurate and efficient zebrafish knockout solutions. With our state-of-the-art CRISPR/Cas9 technology, we ensure precise and reliable genome editing in zebrafish. Our experienced scientists will guide you through the entire process, from sgRNA design to the generation and validation of knockout zebrafish lines.

Why Choose InVivo's Zebrafish Knockout Services?

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.

Knockout Zebrafish Service Options

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

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. 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
  • In silico design of all reagents including the sgRNA site and donor homology template
  • Sequence files for the donor homology construct and edited locus
  • 4 vials of 10uL lyophilized sgRNAs duplexed with the Cas9 protein
  • Injection dyes for early visualization of injection success
  • Detailed Instructions on mix reconstitution and injection protocols.

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?

  • Selection and testing 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 Custom Injection Mix option for the novice zebrafish line builder.

What’s Included?

  • Full in vivo evaluation of somatic editing efficiency.
  • Confirmation of integration capacity of your edit into the zebrafish genome

Mosaic Clutch (F0 injected embryos)

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?

  • Expertly injected embryos for direct submission to you nursery
  • Fully validated PCR primers and protocols for edit detection assay in your own lab.

Full Build (Sequence verified heterozygous line)

Choose this option if you want us to deliver a stable sequence validated CRISPR knockin line.

What’s Included?

  • Growth of F0 injected embryos in our nursery
  • Rigous screening of F0 injected adults for germline transmission of your edit
  • Sequence conformation and propagation of F1 heterozygous animals

Component Descriptions for our Injection Mix & sgRNA Validation Service

More Zebrafish Gene Editing Services

Zebrafish Knockout Service FAQs

Having very high similarity in genetic makeup with humans, zebrafish has proved to be an important model for human genetics and disease. By using zebrafish knockout or knock-in models, genes can be manipulated to understand different disease pathways.

CRISPR/Cas9 Zebrafish Knockout Service is a specialized scientific service that utilizes the CRISPR/Cas9 gene editing technology to create targeted gene knockouts in zebrafish embryos. It enables researchers to precisely disrupt specific genes of interest, providing a valuable tool for studying gene function and understanding the role of genes in various biological processes.

CRISPR/Cas9 Zebrafish Knockout Service works by designing and introducing specific guide RNAs (gRNAs) and Cas9 protein into zebrafish embryos. The gRNAs guide the Cas9 protein to the targeted gene, where it induces double-stranded breaks. These breaks are then repaired by the cell’s natural DNA repair mechanisms, often leading to the introduction of insertions or deletions (indels) that disrupt the function of the targeted gene.

CRISPR/Cas9 Zebrafish Knockout Service offers several advantages. It allows for precise and efficient gene knockout, providing researchers with a valuable tool to study gene function and model human diseases. Zebrafish embryos are transparent, rapidly develop, and share genetic similarities with humans, making them an ideal model organism for genetic research. The service saves time and resources compared to traditional knockout methods and enables researchers to generate multiple knockout lines simultaneously.

Yes, specific genes can be targeted with CRISPR/Cas9 Zebrafish Knockout Service. The design of the gRNAs determines the target gene. Researchers can choose to target any gene of interest by designing gRNAs that are complementary to the desired genomic sequence. This flexibility allows for precise and selective gene knockout.

The timeline for obtaining zebrafish knockout lines using CRISPR/Cas9 Zebrafish Knockout Service can vary depending on various factors, including the complexity of the targeted gene and the efficiency of the gene editing process. Typically, it takes a few weeks to generate founder fish with the desired genetic modifications. The subsequent steps, such as breeding and genotyping, may take several months to establish stable knockout lines.

Contact us so can provide a more accurate timeline based on the specific requirements of the project.

Knockout Zebrafish Publications

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