Infection sensing system with CRISPR in mammalian cell and mammalians

• Project/Area Number: 17K19395
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: Biology of Cells to Organisms, and related fields
• Research Institution: Osaka University
• Principal Investigator: Takeda Junji 大阪大学, 微生物病研究所, 招へい教授 (50163407)
• Project Period (FY): 2017-06-30 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000); Fiscal Year 2018: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2017: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: CRISPR / 感染感知システム / Adaptation / cascade / Cas3 / Cas1 / Cas2 / アダプテーション / 感染検知

Outline of Final Research Achievements

Emergence of CRISPR system has revealed that bacteria also have acquired immunity. When bacteria are infected with phage, part of phage genome is inserted into CRISPR locus in bacteria. This system is called adaptation. In this study, I have tried to reconstruct the adaptation system in mammalian cells. For this, genes involved in adaptation system were codon-optimized and introduced into 293 cells. With detailed analysis by next generation sequencing, I have not detected positive signal for adaptation yet.

Academic Significance and Societal Importance of the Research Achievements

Adaptationシステムは、本来、原核細胞に備わっている外来異物のゲノム情報を自らのCRISPR遺伝子座に取り込むシステムである。そのAdaptationシステムを哺乳細胞で構築する事ができれば、外来病原体の感染履歴をCRISPR遺伝子座を調べる事により、明らかにすることができる。さらに、将来的には同様のCRISPRシステムを有するモデル動物を作製する事により、個体レベルの感染状態をモニターすることが可能になることが期待できる。