Zebrafish Reporter Lines Generation

Due to small size, high fecundity, optical transparency and similarity to humans in genetics and anatomy, zebrafish (Danio rerio) has served as a novel mainstream vertebrate model with significant advantages in studying developmental genetics and human diseases.The larva’s small size allows drug screening to be conducted in multi-well plates, compatible with robotic or channel pipetting, both minimizing compound dosage, and maximizing throughput. The optically translucent larvae, complemented by various fluorescent transgenic strains, promote direct scoring of organ morphology, and cytological events.

There is a growing acceptance that a general biomarker set of genes, which are all equally regulated by various toxicants, does not exist and that toxic compounds regulate common pathways, rather than common genes. This highlights the necessity for the development of an array of reporters and biomarkers. A selection of these biomarkers was used to develop a battery of sensitive and specific fluorescent transgenic zebrafish lines that are able to report cellular toxic load in a dose and temporal specific manner.

Transgenic Zebrafish Reporter Lines

Creative Biogene, a zebrafish research company with experienced scientists and advanced technology, has developed a large number of fluorescent transgenic zebrafish lines and these transgenic zebrafish lines, including gene/enhancer trapped lines, have been targeted for fluorescent protein expression in essentially all tissues and organs.

Figure 1. Microinjection of Cas9 fluorescent fusion proteins into zebrafish embryos. (Burger A, et al. 2016)

• Zebrafish Epitope Tag
  The addition of an epitope tag at the endogenous locus allows you to quantify your protein without altering the level of gene expression. Epitope tags in zebrafish are added through CRISPR/Cas9 gene editing technology. You can choose from a variety of tags including HA, HIS, FLAG, TAP, or S-peptide.
• Zebrafish Fluorescent Tag
  The addition of a fluorescent protein tag at the endogenous locus allows you to visualize your protein in vivo without altering the level of gene expression. In Zebrafish, fluorescent proteins are added by CRISPR/Cas9 gene editing technology. You can choose from various protein tags including GFP, YFP, BFP, and mCherry. Work with our expert genome engineers to choose the tag type and location for knock-in, and we will offer you a customized service package including CRISPR/Cas9-mediated tagging and data curation services. For more information or consultation, please feel free to contact us. Our staff will assist you to get rid of the situation.

References

1. Yao Y,et al. Screening in larval zebrafish reveals tissue-specific distribution of fifteen fluorescent compounds. Disease models & mechanisms, 2017, 10(9): 1155-1164.
2. Zhang X, Gong Z. Fluorescent transgenic zebrafish Tg (nkx2. 2a: mEGFP) provides a highly sensitive monitoring tool for neurotoxins. PloS one, 2013, 8(2).
3. Poon K L, et al. Editor's highlight: transgenic zebrafish reporter lines as alternative in vivo organ toxicity models. Toxicological Sciences , 2017, 156(1): 133-148.
4. Burger A, et al. Maximizing mutagenesis with solubilized CRISPR-Cas9 ribonucleoprotein complexes. Development, 2016, 143(11): 2025-2037.

For research use only. Not intended for any clinical use.