Establishment of highly efficient chloroplast genome editing by using synthetic custom design nucleases

2015 Fiscal Year Final Research Report

• Project/Area Number: 25450002
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Science in genetics and breeding
• Research Institution: The University of Tokushima
• Principal Investigator: Osakabe Keishi 徳島大学, 農工商連携センター, 特任教授 (70450335)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: 葉緑体 / ゲノム編集 / TALEN / Cas9

Outline of Final Research Achievements

To establish chloroplast genome editing in order to open a frontier of chloroplast engineering, site-specific DNA cleavage system by using the synthetic custom design nuclease was designed and constructed. At first, TALEN as a synthetic custom design nuclease was chosen, and The TALEN with transit peptide (tp-TALEN) against to the target on chloroplast genome was constructed. The tp-TALEN could cut the target DNA specifically in vitro, but was failed to transport to chloroplast. Therefore, Cas9 nuclease was used instead of TALEN. Cas9 with transit peptide was successively transported to chloroplast.

Free Research Field

植物分子遺伝学