| title: | The impact of retroposons on the epigenetic regulation of human genes |
|---|---|
| reg no: | ETF5171 |
| project type: | Estonian Science Foundation research grant |
| subject: |
1.6. Physical, Analytical and Inorganic Chemistry |
| status: | completed |
| institution: | Tallinn Technical University |
| head of project: | Mart Speek |
| duration: | 01.01.2002 - 31.12.2005 |
| description: | The importance of ectopic transcription of cellular genes by human L1Hs antisense promoter (ASP) discovered in our laboratory will be studied by the following experimental and in silico approaches: (1) Transcriptional studies involve characterization of the L1Hs ASP (region 400-600 nt), sense promoter (SP) and enhancer element (EE) by using expression of L1Hs ASP/SP/EE deletion and mutation constructs in reporter plasmids, DNase I footprinting in vitro and in vivo, oligonucleotide mobility shifts, determination of transcriptional initiation sites by primer extension and RNAse protection, and identification of transcription factors by South-Western blotting. This study determines promoter (ASP/SP/EE) regions and identifies transcription factors necessary for ASP/SP/EE activity. (2) Search for the potential cellular genes which promoters were captured by L1Hs retrotransposon (graphical Wu-BLAST2, Gene Discovery). Determination of the number and location of possible solo L1Hs promoters in the human genome. Search for the B2 SINE, endogeneous retroviruses and other retroposons involved in the ectopic transcription of cellular genes (EST databank analysis). Predictions made by this section's bioinformatic approach will be verified experimentally using standard molecular biology methods. (3) Transcriptonal interference (promoter occlusion) between L1Hs SP and ASP, and transcriptional termination will be studied by using expression of promoter constructs in dual reporter gene plasmids. (4) Creation and analysis of the transgenic mice expressing L1Hs ASP and cellular gene chimeric constructs. This study reveals the role of L1Hs ASP-induced transcriptional interference or posttranscriptional gene silencing (PTGS) of human/mouse genes. (5) Translational studies of the chimeric L1-cellular mRNAs are used demonstrate the translational potential of these RNAs in vivo.The effects of L1Hs ASP-induced transcriptional interference, promoter insulation, PTGS (epigenetic regulation) and translation of human cellular genes/mRNAs will be determined The effects of L1Hs ASP-induced transcriptional interference, promoter insulation, PTGS (epigenetic regulation) and translation of human cellular genes/mRNAs will be determined. |
| project group | ||||
|---|---|---|---|---|
| no | name | institution | position | |
| 1. | Mart Speek | TTU Center for Gene Technology | senior scientist | |