Using RCAS reporter to analyze the silencing activities of different tissues in chicken embryo
碩士 === 國立中興大學 === 生命科學系所 === 101 === Previous researches showed the transgene expression is hindered in stem and germ cell resulting from the epigenetic silencing. It was also demonstrated that the CpG island methylation silenced viral vector promoter activity. We used RCAS retroviral system to deli...
| Main Authors: | , |
|---|---|
| Other Authors: | |
| Format: | Others |
| Language: | zh-TW |
| Published: |
2013
|
| Online Access: | http://ndltd.ncl.edu.tw/handle/73738016105981579899 |
| Summary: | 碩士 === 國立中興大學 === 生命科學系所 === 101 === Previous researches showed the transgene expression is hindered in stem and germ cell resulting from the epigenetic silencing. It was also demonstrated that the CpG island methylation silenced viral vector promoter activity. We used RCAS retroviral system to deliver exogenous gene into different organs of embryos, and found the expression of these genes showed normal expression in most somatic tissues but very limited in germ cells. We hypotheted that the CpG methylation on the RCAS promoter may have suppresses of gonad-specific expression of RCAS in chicken embryo. We used RCAS-Luciferase to infect chicken embryo, and quantify RCAS promoter activity variation among different tissues. Our result showed RCAS promoter activity in forelimb higher than in gonad tissue. In addition, we used bisulfite sequencing to detect CpG methylation status of the RCAS promoter sequence in those tissues. The sequencing result showed RCAS promoter region express high methylation status in gonad tissue than forelimb.
|
|---|