| Project/Area Number |
23310142
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Applied genomics
|
|---|
| Research Institution | National Institute of Agrobiological Sciences |
|---|
Principal Investigator |
TOKI Seiichi 独立行政法人農業生物資源研究所, ゲノム機能改変研究ユニット, ユニット長 (80212067)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
SAIKA Hiroaki 独立行政法人農業生物資源研究所, ゲノム機能改変研究ユニット, 主任研究員 (20435613)
遠藤 真咲 独立行政法人農業生物資源研究所, ゲノム機能改変研究ユニット, 研究員 (40546371)
|
|---|
| Co-Investigator(Renkei-kenkyūsha) |
ENDO Masaki 独立行政法人農業生物資源研究所, ゲノム機能改変研究ユニット, 研究員 (40546371)
|
|---|
| Project Period (FY) |
2011-04-01 – 2014-03-31
|
| Project Status |
Completed (Fiscal Year 2013)
|
| Budget Amount *help |
¥19,370,000 (Direct Cost: ¥14,900,000、Indirect Cost: ¥4,470,000)
Fiscal Year 2013: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2012: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2011: ¥10,010,000 (Direct Cost: ¥7,700,000、Indirect Cost: ¥2,310,000)
|
|---|
| Keywords | 変異導入 / 人工制限酵素 / 植物 / ゲノム改変 / 非相同末端結合 / 変異スペクトラム / 形質転換 |
|---|
| Research Abstract |
DNA double strand breaks induced by TAL effector nucleases (TALENs) are often imprecisely repaired by non-homologous end joining (NHEJ) and subsequently lead to mutations at repaired site. Here, we investigated whether the lack of DNA ligase 4 (Lig4); one of the key factor of NHEJ pathway ; affected the fidelity and efficiency of NHEJ repair after induction of DSBs by TALENs in rice cells. We found an enhanced mutation frequency in Lig4 mutant.
For establishing an efficient and universal CRISPR/Cas mediated mutagenesis system in rice, we compared different Cas9 expression constructs and found that mutation frequency varied greatly depending on the Cas9 expression constructs. We also revealed that expression level of Cas9, culture period of calli transformed with Cas9 and guide RNA affected mutation frequency.
|
