Zebrafish Tol2 Transgenesis Services

Zebrafish Tol2 Transgenesis Services

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Insert large content in the zebrafish genome using the Tol2 transposon-based system.

Using Tol2 transgenesis you can generate:

  • Cre Recombinase-Driver lines
  • Transcriptional reporters
  • Models to analyze tissue-specific gene expression
  • Models to control the timing of gene expression
  • Humanized zebrafish

Comparison of Tol2 vs. CRISPR

Nature of EditNumber of InsertionsGermline Edits
Germline EfficiencyCommon TerminologyPhenotype in F0 (injected embryos)Max. Size of Insert
Tol2 Transposase
Random insertion
Multiple
Yes
10% to 40%
Transgenesis
Mosaic
Usually 13-15kb - depends on cargo
CRISPR/Cas9
Site specific
Single
Yes
~2% to 5%
Knock-in Edit
Mosaic
Unknown but appears that the larger the insert the lower the efficiency.

Tol2 Transgenesis Service Options (price reflects academic pricing)

Standard Injection Mix# Complete Validated Injection Mix Verified Clutch* (F0 injected Embryos) Full Build** (Sequence verified heterozygous line)
Tol2 Plasmid Design
Injection Mix Assembly
InVivo Integration Assessment
Expertly Injected Embryos
Germline Transmission
Screening & Line Propagation
Pricing*$2,995$5,595$7,595$21,840
Timeline4-6 weeks4-8 weeks1-2 months4-6 months
More DetailsMore DetailsMore DetailsMore Details
* This service is only available to clients in the United States.
** Full Build includes screening n=100 adults; standard husbandry charge applies.
Additional charge applies for cargo over 2kb in size for all service packages.

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.

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

Verified 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

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 as a stable dehydrated reagent.

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

  • Custom reagents from validated suppliers
  • Four 10uL lyophilized injection mixes with instructions on reconstitution
  • Rhodamine dye for early visualization of injection success
InVivo Integration Assessment

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
Expertly Injected Embryos

Expertly Injected Embryos

Our team precisely injects the sg(s) targeting the locus or region of interest, Cas9 protein and the donor homology template to generate F0 embryos.

Germline Transmission Screening & Line Propagation

Germline Transmission Screening & Line Propagation

F0 embryos are reared to adulthood and then screened for germline transmission using a combination of screening techniques to identify germline founders. Identified adults are propagated to ensure a sufficient number of animals to ship to your facility.

READY TO GET STARTED?

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

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