Zebrafish embryo microinjections are used for manipulating gene expression, making targeted genome edits and generating transgenic lines.

mRNA and genomic targeting

• Transient mRNA "knock-downs" with anti-sense morpholinos
• Stable mutant fish lines (CRISPR-Cas9)

Overexpression

• Transient overexpression using 5’-capped polyadenylated mRNAs
• Stable overexpression with transgenic lines

Transgenic reporters

• Transient gene reporter assays
• Stable transgenic reporter gene lines

Microinjection is offered on a fee-for-service basis. The user provides the necessary reagents; the facility provides the embryos and carries out injections. Samples and work-up are discussed prior to the experiments. The platform can also provide help for certain technical aspects of an experimental design. For example, this could be help in designing a zebrafish CRISPR-Cas9-based gene targeting strategy, a transgenesis construction, a genotyping approach or zebrafish-related protocols.

This is charged at 150 CHF for a half day of experimentation.

If stable fish lines are to be generated there is a charge for housing the fish per tank and per day (see Housing zebrafish lines).

If a user already has experience with the zebrafish model and would like to have access to the micro-injection setup, the same 150 CHF fee is charged for a half day access. Fish crosses and previous experience with this equipment need to be discussed in advance.

The core facility can provide zebrafish embryos for direct experimentation, up to day five of development, as well as developmentally staged embryos which are either paraformaldehyde-fixed or in Trizol. Several laboratory zebrafish backgrounds are available.

Each sample is charged at 50 CHF, typically as a pool of 50 embryos.

The core facility can house zebrafish lines for users. Each aquarium used for this service is charged at 30 CHF per month. For example, a novel zebrafish mutant (one allele) generated using CRISPR-Cas9 requires 3-4 tanks for 3 generations of fish (3-4 tanks for 9-12 months = 810 to 1440 CHF).

The core facility has a ViewPoint swimming behavior instrument. It measures swimming speed and quality and can stimulate larvae with light or vibrations while monitoring. Use of the instrument is charged at 25 CHF per hour.

The core facility has a biosafety level 2 laboratory for studies of microbial infection and inhibition. For example, we can microinject fluorescent bacteria into zebrafish embryos and then treat with potential anti-microbials. Fluorescence can be monitored and bacteria quantified over time using instrumentation in the READS platform. To date this has been used for studies of M. marinum infectivity and inhibition.

The platform can provide a limited number of standard reagents and consumables. For example, this will include embryo media, plastics, control morpholinos, additives for micro-injections and recombinant Cas9. Any minor additional cost to users for such reagents will be discussed prior to experimentation.

Discussions on planning an experiment are free of charge. This may include an assessment of feasibility, planning a time-frame for experiments and their overall costing including specific reagents.

Detailed experimental design is charged at 150 CHF per two hours of working time. For example, users can ask for data mining for information on specific gene orthologs in zebrafish, or help in the design of a CRISPR-Cas9 mutagenesis approach for their gene-of-interest. Detailed help with veterinary authorizations is also charged at this rate.

Microinjection is charged at 150 CHF per session. This is typically fish mating setup and 2 hours of injections.

Imaging is charged at 150 CHF for each 2 hours.

Zebrafish samples are charged at 50 CHF per sample, this includes the provision of embryos.

Use of the biosafety level 2 laboratory is charged as for regular experimentation.

Zebrafish housing is charged at 30 CHF per month per aquarium. An aquarium holds up to 17 adult fish.

A small fee for general reagents are added to the above prices. This covers buffers, plasticware and simple solutions used in microinjections and is typically 50 CHF per experiment.

Behavioral analysis is charged at 25 CHF per hour.

Project-specific reagents are payed for by the user. For example, morpholino anti-sense oligonucleotides, mRNAs, gRNAs, etc.