Below is a listing of our services and techniques.

Services

You can use our expertise to model your disease of interest in zebrafish. The core can plan, design, and analyze a disease model for you. Typically, we create a gene knock out or knock-in, analyze the phenotype, and help to compare the results to a patient’s disease. As next steps we can analyze disease mechanisms in detail and start a drug screen to find reagents that can overcome a disease state.

We train researchers to become zebrafish experts. We provide training in all fish-relevant techniques, help you to get a project started, and provide consultations during ongoing projects.

We take care of your fish and grow the larvae, and ensure the water parameters are where zebrafish like them.

The Zebrafish Core has a single IACUC protocol in which all important zebrafish techniques are described. New projects can be easily added on by a brief summary.

Imaging is an essential part of zebrafish research. The larvae are clear and all organs are visible. The size of a 5-day-old larvae is ideal for microscopy; it is thin enough to focus through a whole larvae, and organs are small enough so an entire organ can be imaged with a confocal microscope and a 20x lens. Zebrafish researchers have created fluorescent lines labeling nearly all organs or specific tissues within organs. Time-lapse analysis is another hallmark of zebrafish research, allowing fish to be followed under a microscope for hours or days so observation of organ development, nerve growth, tissue interaction during tumor formation, or whole animal development in mutants is possible.

Most agents can be injected in the first forming cell with a fine glass needle and are then passed along to the dividing cells. CRISPR/Cas9 knock-outs are efficient and the core can create a complete knock out of a locus in the injected generation for most genes. Transgenic expression and integration is also efficient due to special tools. Most zebrafish researchers use the gateway cloning system that relies on 3 fragments (e.g. promoter, gene, fluorescent tag) that is cloned in a transgenesis vector; the fragments are interchangeable and a large tool box of different promoters, genes, and labels is available.

The accessibility to gene knockouts and transgenes enables zebrafish to serve as a disease model so investigators can screen for drugs and the results can translate to the clinic. Fish can be grown in small groups in 6- to 96-well pates. Drugs can be added to the water and most are taken up through the skin, intestine, or thin gill epithelium. Due to the clarity and size of larvae, drug rescue is easily detectable.

Zebrafish have a set of defined behavioral patterns that can be used for tests. The Zebrafish Core can monitor activity (e.g., speed, velocity, frequency) and response to specified stimuli.

Techniques

  • CRISPS/Cas9 knockouts
  • Morpholino gene knock down
  • mRNA injection
  • Tilling lines (from Sanger Center/ZIRC)
  • Expression constructs for transgenic lines
  • Creation of transgenic lines
  • Genotyping
  • Help with analytic assays (Microarray, RNAseq.)
  • Confocal imaging
  • Imaging of transgenic proteins (GFP, mCherry)
  • Histological thin sectioning of larvae
  • Gene and protein expression analysis (antibody, mRNA in-situ hybridization
  • WT eggs for injection
  • Mutant and transgenic larvae (as requested)
  • Animal husbandry