Establishment of a new method for functional analysis of cis-regulatory modules using CRISPRi

2016 Fiscal Year Final Research Report

• Project/Area Number: 26650087
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Developmental biology
• Research Institution: Okinawa Institute of Science and Technology Graduate University
• Principal Investigator: Yasuoka Yuuri 沖縄科学技術大学院大学, マリンゲノミックスユニット, 研究員 (70724954)
• Research Collaborator: TAIRA Masanori 東京大学, 大学院理学系研究科生物科学専攻, 准教授 (60150083)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: ゲノム編集 / 転写制御 / ツメガエル

Outline of Final Research Achievements

It is important to analyze in vivo functions of cis-regulatory modules during embryogenesis. However, quick and effective methods for such analysis have never been established. Therefore, I examined whether CRISPRi by a dead Cas9 protein (dCas9) can regulate activities of cis-regulatory modules using Xenopus embryos. Luciferase reporter assays were performed using both dCas9 mRNA and protein. The data showed that CRISPRi could not affect the reporter gene expression. This result suggests that endogenous transcriptional machineries can bump CRISPRi/dCas9 from DNA.

Free Research Field

発生生物学・進化生物学