Molecular Mechanism of Early Retinal Development by the RNA-binding Protein Quaking

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K09839
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 56060:Ophthalmology-related
• Research Institution: Ritsumeikan University
• Principal Investigator: Moritoh Satoru 立命館大学, 薬学部, 助教 (20647234)
• Co-Investigator(Kenkyū-buntansha): 小池 千恵子 立命館大学, 薬学部, 教授 (80342723) ; 川村 晃久 立命館大学, 生命科学部, 教授 (90393199)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: マウス / 網膜 / 3次元網膜 / Quaking / コンディショナルノックアウト

Outline of Final Research Achievements

To analyze the function of the RNA-binding protein Quaking (Qki) in the retina, we generated Qki-disrupted ES cell lines by the CRISPR/Cas9 method and differentiated them into 3D retinal cells. Rx-GFP positive retinal progenitor cells differentiated and an optic vesicle-like structure was observed, unlike in the Qki knockdown experiment. These results indicate that, contrary to initial expectations, Qki is unlikely to be involved in early retinal developmental stages such as retinal progenitor cell formation and optic vesicle formation. Therefore, for functional analysis at the in vivo level, retina-specific Qki conditional knockout (CKO) mice were generated by crossing Qki flox mice with retina-specific　Cre mice, and histological analysis by immunostaining revealed that Qki was knocked out in the retina-specific manner. We are currently conducting a more detailed analysis.

Free Research Field

眼科学

Academic Significance and Societal Importance of the Research Achievements

本研究でのRNA結合タンパク質Quakingに着目した網膜発生の個体レベルでの機能解析により、網膜発生や分化の理解が深まることで、将来的に網膜分化系の改良など再生医療や創薬のための重要な基礎研究となることが期待される。