Development of applicable methods for one-step generation of genetically modified mammals specialized in brain function analysis

2015 Fiscal Year Final Research Report

• Project/Area Number: 26640007
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Neurophysiology / General neuroscience
• Research Institution: Niigata University
• Principal Investigator: Sakimura Kenji 新潟大学, 脳研究所, 教授 (40162325)
• Co-Investigator(Renkei-kenkyūsha): Abe Manabu 新潟大学, 脳研究所, 准教授 (10334674)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: 遺伝子編集 / CRISPR/Casシステム / 胚操作 / コンディショナルノックアウト / レンチウイルスベクター

Outline of Final Research Achievements

The main aim of this research is to develop basic methods for one-step generation of genetically modified mammals. For this purpose, I had planned to develop new methods using the CRISPR/Cas system for generation of genetically modified mice equivalent to conditional knockout mice.
I created two mutant mouse lines which could be induced to express Cas9 proteins depending on the activity of Cre recombinase or tetracycline-transactivator. To introduce a foreign gene into embryos, I confirmed a technique for efficient gene transfer using a lentiviral vector. Furthermore, I demonstrated that efficient one-step insertion of a short tag was preformed by electroporation using embryos. Because these methods could be adapted to other mammalian species in principle, we believe that these will contribute to analyzing the molecular mechanisms of higher brain functions and diseases as basic technologies using gene modified animals such as monkey.

Free Research Field

分子神経生物学