Development of Y chromosome multicopy gene knockdown mice using CRISPR interference

Research Project

Project/Area Number	16K14596
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Multi-year Fund
Research Field	Laboratory animal science
Research Institution	Japan Advanced Institute of Science and Technology (2017) The University of Tokushima (2016)
Principal Investigator	SAKARI MATOMO 北陸先端科学技術大学院大学, 先端科学技術研究科, 研究員 (90466772)
Co-Investigator(Renkei-kenkyūsha)	MATSUMOTO TAKAHIRO 徳島大学, 大学院・動物資源研究部門, 准教授 (70447374)
Research Collaborator	ATSUMI KENGO 徳島大学, 医学部 OHNO HITOSHI 徳島大学, 医学部, 研究協力員 AKAZAWA EMIKO 徳島大学, 医学部, 研究協力員
Project Period (FY)	2016-04-01 – 2018-03-31
Project Status	Completed (Fiscal Year 2017)
Budget Amount *help	¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000) Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000) Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywords	Y染色体 / 精子形成 / 多コピー遺伝子 / CRISPR干渉法 / 応用動物
Outline of Final Research Achievements	This study aims to develop Y chromosome multicopy gene knockdown mice using CRISPR interference, development of knockdown mice using artificial miRNA, development of knockdown mice using CRISPR interference, reproduction We aim to elucidate the cell-specific splicing mechanism. We succeeded in identifying the splicing target genes by continuous generation of knockdown mice as the result of the whole period and by next generation sequencer analysis using germ cell line. Furthermore, transcriptional analysis of the target genes revealed a novel transcriptional regulatory mechanism that negatively regulates the RNA polymerase's pausing mechanism. These results suggest that pausing control of RNA polymerase is an important mechanism in spermatogenesis process.