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CRISPR/Cas9 Zebrafish Knockout Services for Precise Genome Editing

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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.

Table of Contents

Knockout Zebrafish Service Options

Standard Injection Mix

Validated sgRNA Injection Mix

Fully Validated Injection Mix

Mosaic Clutch* (F0 injected embryos)

Full Build** (Sequence verified heterozygous 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?

  • OUR VALIDATED sgRNA INJECTION MIX PLUS:
  • 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?

  • OUR VALIDATED sgRNA INJECTION MIX PLUS:
  • 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?

  • OUR COMPLETE VALIDATED INJECTION MIX PLUS:
  • 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?

  • VERIED CLUTCH (F0 INJECTED EMBRYOS) PLUS:
  • 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

Injection Mix Service Component Descriptions

More Zebrafish Gene Editing Services

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Zebrafish Injection Mix


Knockout Zebrafish Publications

Resources

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