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Policies & Fees
Division of Labor
Because of limited staff, the core favors a team-based approach for more involved projects. A team-based approach means that the core will provide training, access to equipment and standard reagents and quality-control feedback to someone from the recipient laboratory who will independently perform key parts of the needed experiments. This policy is essential to ensure that the core can serve everybody's needs. Shorter projects requiring just a few rounds of a single procedure, or work with regulated animals in the case of non-licensed researchers, will be performed exclusively by the core's staff.
Good core citizenship means reserving and logging the time that you use core equipment, proper use of the equipment and being mindful of others in the core's shared space. To gain access, please return a signed copy of the NICHD Zebrafish Core User Agreement to Ben Feldman.
Charges for Service and Consumables
Core costs and pricing are still being evaluated. At present, consultation and training time, common reagents and instrument use are all free of charge but laboratories will be asked to pay for costly consumables.
Credit for Published Work
To help ensure our continued funding, it is critical that recognition be given on publications that include data generated with the core's assistance. Whether this recognition comes as co-authorship for a core staff member or as an acknowledgement should depend upon the significance of the core's contribution.
Equipment & Reagents
Equipment maintained by the core:
- Dissection microscope and microinjection station
- Eco RT-PCR machine
- Semi-automatic in situ hybridization machine
Equipment maintained by the core and shared with NHGRI:
- Live imaging station with fluorescent and time-lapse capacities
- Compound upright Zeiss microscope
Also maintained by the core:
- List of protocols
- Toolkit for TALEN assembly
- DNA vectors and constructs for RNA synthesis and transgenesis
Ancilllary equipment maintained by NICHD:
- Bulk zebrafish embryo spawning system
- Liquid nitrogen storage for frozen zebrafish sperm
Links & Databases
Please contact the core with new link suggestions or questions about how to use these resources.
Main Zebrafish Research Community Site
ZFIN, The University of Oregon's Zebrafish Information Network
Washington Area Zebrafish Labs
Zebrafish Interest Group
Ensembl's Zebrafish Genome Browser
UCSC's Zebrafish Genome Browser
Finding Zebrafish Lines
ZFIN's General Fish Database
The University of Oregon's Zebrafish International Resource Center Homepage & General Fish Database
The Sanger Institute's Zebrafish Mutation Project Homepage & Fish Knockout Database
The Fred Hutchinson Cancer Research Center Moens TILLing Project Homepage & Fish Knockout Database
The National Institute of Genetics Kawakami Lab's Transgenic Fish & Fish Knockout Databases
The Institute of Molecular and Cell Biology's Zebrafish Enhancer TRAP lines database Homepage & Transgenic Fish Database
Center for Regenerative Therapies Dresden's CreZoo Transgenic Fish Database
TALENs in Zebrafish
Zebrafish mutants vs. mouse mutants: a comparison
The case for zebrafish mutants
- Zebrafish are evolutionarily closer to humans than most people realize. Indeed, after mice zebrafish is the next closest standard genetic model organism.
- TALENs are an exciting new technology, and the procedure for generating pairs of TALENs that can mutagenize your zebrafish locus of interest is quite fast.
- The rapid and ex vivo nature of zebrafish embryogenesis as well as their optical transparency can greatly simplify the analysis of embryonic phenotypes relative to mice.
- Zebrafish can produce hundreds of embryos in a single mating, allowing for the design of experiments involving many more conditions and more robust statistics than are typically conceivable with mice.
- Zebrafish housing costs are much lower than mouse housing costs.
The case for mouse mutants
- Pre-mutagenized embryonic stem cell lines are available for most mouse genes.
- Mice are evolutionarily closer to humans than zebrafish, making mice frequently the model of choice for biomedical questions.
- Zebrafish research is not faster in terms of breeding and generation time. In fact, the generation time (fertilization to sexual maturity) for zebrafish and mice are very similar: about 3 months. For this reason once a germ-line transmitting chimera is created in either species, an additional 6 months is required before recessive homozygous embryos will be produced.
- Zebrafish are not really faster for detailed phenotype analysis, since ultimately the same set of questions at the same level of detail using the same general methods will be required for a good publishable study.
Still interested in making a zebrafish mutant?
Below is an overview of the steps we undertake for TALEN-based mutagenesis in the NICHD zebrafish core. Several specific websites are mentioned in these pages. There are certainly alternate strategies, websites and web-based tools that can achieve the same goals. In no way do we wish to advocate one of these over another. These are simply the approaches that the NICHD Zebrafish Core currently employs. Researchers interested in minimizing efforts should be aware that several companies offer to design and synthesize TALENs for a fee.
Finding zebrafish orthologs to genes from other species
Deciding where to make the mutation