Gene knockdown by morpholino-modified oligonucleotides in the zebrafish (Danio rerio) model: applications for developmental toxicology.
The zebrafish (Danio rerio) has long been used as a model for developmental biology, making it an excellent model to use also in developmental toxicology. The many advantages of zebrafish include their small size, prolific spawning, rapid development, and transparent embryos. They can be easily manipulated genetically through the use of transgenic technology and gene knockdown via morpholino-modified antisense oligonucleotides (MOs). Knocking down specific genes to assess their role in the response to toxicant exposure provides a way to further our knowledge of how developmental toxicants work on a molecular and mechanistic level while establishing a relationship between these molecular events and morphological, behavioral, and/or physiological effects (i.e., phenotypic anchoring). In this chapter, we address important considerations for using MOs to study developmental toxicology in zebrafish embryos and provide a protocol for their use.
Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA, USA. atimmelaragy@whoi.edu
Alicia R
AR
Timme-Laragy
Sibel I
SI
Karchner
Mark E
ME
Hahn
D000818:Animals; D002138:Calibration; D046508:Culture Techniques; D004195:Disease Models, Animal; D047108:Embryonic Development / Q000235:genetics; D055785:Gene Knockdown Techniques; D005818:Genetic Engineering / Q000295:instrumentation / Q000379:methods; D008845:Microinjections / Q000295:instrumentation / Q000379:methods; D060172:Morpholinos / Q000235:genetics*; D009339:Needles; D011078:Polychlorinated Biphenyls / Q000633:toxicity; D059386:Reverse Genetics; D013723:Teratogens / Q000633:toxicity; D018675:Toxicity Tests / Q000295:instrumentation / Q000379:methods; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*; D029961:Zebrafish Proteins / Q000235:genetics
D060172:Morpholinos; D013723:Teratogens; D029961:Zebrafish Proteins; D011078:Polychlorinated Biphenyls; C023035:3,4,5,3',4'-pentachlorobiphenyl