Development of gene repression system using TALE in the silkworm

Research Project

Project/Area Number	16K18826
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Multi-year Fund
Research Field	Insect science
Research Institution	National Agriculture and Food Research Organization
Principal Investigator	TSUBOTA Takuya 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, 主任研究員 (00612772)
Project Period (FY)	2016-04-01 – 2019-03-31
Project Status	Completed (Fiscal Year 2018)
Budget Amount *help	¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000) Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Keywords	ゲノム編集 / TALEN / カイコ / 遺伝子組換え / 遺伝子発現制御 / TALE
Outline of Final Research Achievements	Constructs encoding TALERs (Fusion protein of TALE and transcriptional repressor domain) targeting silkworm h-fibroin (h-fib) and its repressor Arrowhead (Awh) were generated, and they were microinjected into the silkworm embryo to establish transgenic strains. For TALER of h-fib we could establish the transgenic strain, and this strain was crossed with GAL4 strain for the overexpression of TALER in the silk gland. However, the transcriptional repression of h-fib gene was not observed.
Academic Significance and Societal Importance of the Research Achievements	カイコではRNAiによる遺伝子ノックダウンを行うことが著しく困難であり、その代替技術としてTALENの制限酵素部位FokIと転写抑制因子のリプレッサードメインとを差し替えたTALERタンパク質の利用を試みた。本研究では標的遺伝子の発現抑制を確認することはできなかったが、今後改良を加えていく上での足がかりとなる知見を得ることができた。