| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Outline of Final Research Achievements |
We developed a simple method to use the Cre-loxP recombination system for marker gene rescue in A. oryzae by direct introduction of Cre into protoplasts using DNA as a carrier for the enzyme. To apply the simple method to artificial nucleases (TALEN) for genome editing, we examined kinds of the carrier and conditions of Cre without the carrier for direct introduction. In the carrier, RNA and heparin as well as DNA were available for the carrier. We showed that to raise the electric charge and the concentration of Cre was available for the direct introduction without using the carrier. In addition, because the genome editing of A. oryzae via the error of nonhomologous end-joining repair by a transient expression of TALENs was confirmed, the recombinant TALENs were purified. Hence, we will perform the direct introduction for the genome editing of A. oryzae using the purified TALENs.
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