Establishing a transcriptional regulation system of the p21 gene by artificial transcription factors using genomic editing technique

2014 Fiscal Year Final Research Report

• Project/Area Number: 25550029
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Osaka University
• Principal Investigator: FUJIWARA Tomoko 大阪大学, 医学(系)研究科(研究院), 助教 (70402922)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: メダカ / ゲノム編集技術 / 転写制御 / p21 / 放射線 / 損傷応答

Outline of Final Research Achievements

Recently genome editing techniques such as TALENs and CRISPR has been reported as attractive novel gene targeting method. Both methods include sequence specific DNA binding, and subsequent introduction of DNA double strand break in vicinity of the binding site. In this study, we tried to establish a transcriptional regulation system in Medaka. We generated the Artificial Transcription Factor (ATF) by replacing the DNA cleavage activity with the transcriptional regulation activity. For this purpose we established in vitro transcription regulation system in β-actin and p21 genes. guideRNA were designed at several positions of upstream in each gene. These gRNA and nuclease-null Cas9 construct which tethering transcriptional activation domain include were introduced in medaka cells, and these transcriptional activation activity were detected as luciferase reporter activity. We detected gRNA mediated transcriptional activity in β-actin and p21 gene by this ATF system.

Free Research Field

放射線・化学物質影響化学