CRISPR/Cas9 in zebrafish (Danio rerio): standardization and optimization to identify mutants

• Grant number: 19/15594-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Effective date (Start): September 01, 2019
• Effective date (End): October 31, 2020
• Field of knowledge: Agronomical Sciences - Fishery Resources and Fishery Engineering - Aquaculture
• Principal Investigator: Rafael Henrique Nóbrega
• Grantee: Jéssica Yumi Yamamoto Kita
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Genome editing techniques have allowed researchers to expand knowledge about biological systems mechanisms, as well as, to understand the influence and function of different genes in the whole body. The newest gene editing technique is CRISPR / Cas9 system, that perform mutations more accurately and efficiently due to its simplicity regarding the use of a guide RNA that will aid the Cas9 endonuclease to cleave the target sequence. However, studies involving this technique could be improved to identify the mutated individuals. In addition, a vertebrate animal research model highlighted for its genetic similarity to humans is zebrafish (Danio rerio). Zebrafish has interesting characteristics related to laboratory researches due to its size, high fertilization rate, ex vivo fertilization and embryogenesis, body transparency in early stages of development, rapid embryological development, low maintenance cost and known genome. Therefore, it is essential to improve the genome editing techniques in zebrafish, so that, by increasing the mutation efficiency rate, it is possible to create an optimized method for identifying mutated animals, so it will be possible to extend and improve future studies on zebrafish in medical areas and so on.