Knockout Zebrafish

Zebrafish, a tropical freshwater fish, is originally from the Himalayas. Its body is silver or gold, and there are five stripes on each side. These stripes extend from the head to the back of the caudal fin. Similar stripes are found on the buttocks and caudal fins. These stripes look similar to the zebra stripes, that is why it called Zebrafish. Zebrafish is a popular ornamental fish, and it is also an important and widely used model animal in the field of scientific research. Especially in the research of regeneration, there are many genetically modified varieties.

Zebrafish

Different Stages of Zebrafish

Zebrafish, as a model animal, has the following advantages:

1. Zebrafish experiment is a holistic animal study. Compared with cell experiment in vitro, zebrafish can simulate more complex biological processes.

2. Zebrafish has small size, strong reproductive capacity and large spawn quantity, which is conducive to statistical analysis and suitable for high-throughput research.

3. The whole genome sequencing of zebrafish has been completed, and it has high similarity with human genes, and its gene editing methods have been very mature.

4. Zebrafish embryos is large, prominent and transparent. Its embryos develop rapidly in vivo and in vitro. They are very easy to observe.

5. Zebrafish project has short turnaround and low cost.

6. The ethical risk of zebrafish experiment is low, so the research can be carried out quickly.

※Ubigene provides high quality zebrafish gene knockout services and transgenic services.

1. Technical Details：

Ubigene uses CRISPR/Cas9 method to edit Zebrafish’s genome. CRISPR/Cas9 system is a versatile tool for gene modification of nearly all types of cells. crRNA (CRISPR-derived RNA) binds with tracr-RNA (trans-activating RNA) by base pairing to form tracrRNA/crRNA complex, which guides the Cas9 nuclease to cleave and introduce sequence-specific double stranded DNA breaks (DSB). By designing tracrRNA and crRNA, we can transform them into sgRNA, which is a synthetic RNA sequence that could guide Cas9 to bind and splice the targeted sequence of DNA.

Cas9 nuclease has two domains, NHN (cutting the complementary strand of crRNA) and RuvC (cutting the non-complementary strand of crRNA), which could cleave two single strands of DNA respectively and result in double stranded DNA breaks (DSB). To repair the DSB, the cell uses its own DNA repair machinery to add or delete or replace pieces of DNA sequence via Homology Directed Repair (HDR) or Non-Homologous End Joining (NHEJ).

Repair of DNA Double-strand Break