Development of highly efficient and accumulated protein expression system targeted to fish eggs

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K14984
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Aquatic life science
• Research Institution: Mie University
• Principal Investigator: Tamaru Yutaka 三重大学, 生物資源学研究科, 教授 (50324554)
• Research Collaborator: Nakatani Hajime 名古屋大学, 工学研究科, 講師 ; Hori Katsutoshi 名古屋大学, 工学研究科, 教授
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: ゲノム編集技術 / ゼブラフィッシュ / Crispr-Cas9

Outline of Final Research Achievements

We attempted to construct a functional plasmid for CRISPR-Cas9 system in zebrafish. While the EF-1 alpha (zef1alpha) promoter from zebrafish developed by our laboratory was used for Cas9 protein expression, the U6 promoter newly cloned from zebrafish was used for the expression of guide RNA (gRNA). After an injection to fertilized eggs from zebrafish with the constructed plasmid, both EGFP expression downstream of Cas9 gene in the plasmid and Cas9 protein expression were observed among zebrafish 24 hours-per-fertilization (hpf) embryos. Furthermore, gRNAs designed by the vitellogenin gene in zebrafish genome were successfully expressed.

Free Research Field

水圏生命科学