| Project/Area Number |
15510161
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
基礎ゲノム科学
|
|---|
| Research Institution | RIKEN (The Institute of Physical and Chemical Research) |
|---|
Principal Investigator |
KIYOSAWA Hidenori RIKEN, Technology and Development Team for Mammalian Cellular Dynamics, Research & Development Scientist, 動物変異動態解析技術開発チーム, 開発研究員 (30295422)
|
|---|
| Project Period (FY) |
2003 – 2004
|
| Project Status |
Completed (Fiscal Year 2004)
|
| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2004: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2003: ¥2,000,000 (Direct Cost: ¥2,000,000)
|
|---|
| Keywords | antisense RNA / non-coding RNA / gene expression / microarray / マイクロアレイチップ |
|---|
| Research Abstract |
I had preliminary data of 2500 pairs of sense-antisense genes identified in silico. These pairs of the genes were identified using cDNA data, but direct evidence of their expression had not been obtained. In order to confirm the actual expression of these genes, microarrays (60mer oligo nucleotides) that enabled us to distinguish sense and antisense gene expression and quantitation were constructed. Out of 2500 pairs above,2000 pairs were chosen and subject to expression analysis in fibroblast cells, brain, heart, and testis. It turned out that more than 90% of the genes were actually transcribed, and that the expression valance between the sense and antisense genes varied among tissues.
Seven gene pairs were subject to Northern analysis. Various-sized transcripts were detected as smear on the blots, regardless of coding or non-coding status of the genes. They were mostly poly(A)-negative and tended to be localized in nucleus. These finding suggested that it was difficult to knockout or knockdown the antisense genes only to continue functional analysis by conventional methods.
I analyzed seven sense or antisense genes on X chromosome whether or not they escaped from X chromosome inactivation. Their expression was under X chromosome inactivation.
We found out unusual characters of the RNA transcribed from the sense-antisense gene loci. In order to further analyze function of these transcripts, it is necessary to precisely identify the transcription unit in each locus.
|
