Visualization of the RIP machinery in living Neurospora

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K14916
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied molecular and cellular biology
• Research Institution: University of Fukui
• Principal Investigator: Honda Shinji 福井大学, 学術研究院医学系部門, 助教 (90632167)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: ゲノム防御 / ゲノム編集

Outline of Final Research Achievements

The filamentous fungi Neurospora crassa has a unique genome defense system called Repeat Induced Point mutation (RIP). During specific sexual phase, RIP detects duplicated sequences throughout genome and mutates them, ultimately resulting in gene disruption. The mechanism, however, remains almost unknown. This study aimed to label artificial duplicated sequences by fluorescent proteins and to trace the RIP machinery in living cells. We first optimized blue, green, red, and near-infrared fluorescent protein tagging systems and successfully visualized distinct nuclear compartments in Neurospora. Furthermore, we established a method to effectively produce transparent perithecia to enhance the detection of fluorescent proteins in sexual phases. We are currently optimizing the visualization of artificial duplicated sequences tethered by our improved fluorescent protein systems.

Free Research Field

エピジェネティクス