Technology

Zebrafish CRISPR Knock-Out (KO) Service

Promotor, regulatory elements and exon deletions can be ordered at an extra cost.


Package	Target Validation Basic	Target Validation Plus	Injection Basic (F0 mosaic embryos)	Injection Plus (F0 mosaic embryos)	Full Package Basic (Sequencing verified F1 heterozygotes)	Full Package Plus (Verified F2 mutant homozygotes)
sgRNA & sequencing primers design	+	+	+	+	+	+
In vivo sgRNA activity testing		+	+	+	+	+
Embryos injection			+	+	+	+
Line propagation and mutation screening					+	+
Morphometric evaluation of the phenotype				+	+	+

Zebrafish CRISPR Knock-In Service

Standard service includes insertion of the GFP protein under the expression of the endogenous gene promoter. Other methods can be applied at the customized pricing model.


Package	Target Validation Basic	Target Validation Plus	Injection Basic (F0 mosaic embryos)	Full Package (Sequencing verified F1 heterozygotes)
sgRNA & sequencing primers design	+	+	+	+
In vivo sgRNA activity testing		+	+	+
Embryos injection			+	+
Line propagation and mutation screening				+

Live imaging

Life Imaging with high throughput automated VAST system (Union Biometrica)

Specification

Embryo and larvae 3-6 days post fertilization, 96-well plate based, bright field and fluorescent (green and red) imaging, magnification: 4X, 5X, 10X, (20X), automated positioning of the fish, image analysis option

Flat rate of 1000 sek/run + 250 sek/h of the run

Fluorescent microscope image of zebrafish trunk

Toxicity tests

Fish embryo acute toxicity (FET)

Test based on the adapted Test Guideline 236. This test is designed to determine acute toxicity of chemicals on embryonic stages of fish. Recommended by the EU grant committee as an intermediate step between in vitro and in vivo mouse studies.

Cancer modelling

STEP 1 – Evaluation of the human cancer cell line xenografts in the zebrafish embryo

An assay designed to evaluate cancer cell line compatibility with zebrafish model, including cell size, behavior, survival, engraftment side and embryo toxicity.

Cell survival was tested in vivo at 33°C
Cells are fluorescently labeled
Cells are provided in 0.25-0.5x106 cells/ul concentration

STEP 2 – Tumour growth & metastasis assay

Customized experimental set up based on the results of the evaluation assay, injections at 1 or 2 dpf into the blood flow, brain cavity or PVC space. Imaging of the xenotransplanted fish at 2 time points. Possibility of combining xenotransplantation procedure with the drug treatment