Application of the gene knockout method using CRISPR / Cas system to the hemipteran insect

Research Project

Project/Area Number	26660274
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Multi-year Fund
Research Field	Insect science
Research Institution	National Agriculture and Food Research Organization (2016) National Institute of Agrobiological Sciences (2014-2015)
Principal Investigator	Matsumoto Yukiko 国立研究開発法人農業・食品産業技術総合研究機構, 生物機能利用研究部門昆虫制御研究領域, 主任研究員 (80414944)
Co-Investigator(Renkei-kenkyūsha)	HATTORI Makoto 独立行政法人農業生物資源研究所, 加害・耐虫機構研究ユニット, 研究専門員 (60370673)
Project Period (FY)	2014-04-01 – 2017-03-31
Project Status	Completed (Fiscal Year 2016)
Budget Amount *help	¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000) Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Keywords	CRISPR / ツマグロヨコバイ / Nephotettix cincticeps / Parental RNAi / parental RNAi
Outline of Final Research Achievements	I tried CRISPR / Cas 9 in Nephotettix cincticeps (Hemiptera: Cicadellidae). RNA was injected into adult females, and genome modification in the next generation larvae was expected. It is the same method as parental RNAi, therefore, parental RNAi method was confirmed to be effective in N. cincticeps ( Arch Insect Biochem Physiol. 91:152-164.). CRISPR / Cas9 was performed on female adults using NcLac1S (a salivary gene) and NcLac2 (an epidermal laccase gene), and the genomic DNA of the next generation larvae was checked, but unfortunately no deletion or modification was observed.