Abstract
The exact mechanisms leading to alternative splice site selection are still poorly understood. However, recently cotransfection studies in eukaryotic cells were successfully used to decipher contributions of RNA elements (cis-factors), their interacting protein components (trans-factors) or the cell type to alternative pre-mRNA splicing. Splice factors often work in a concentration dependent manner, resulting in a gradual change of alternative splicing patterns of a minigene when the amount of a trans-acting protein is increased by cotransfections. Here, we give a detailed description of this technique that allows analysis of large gene fragments (up to 10-12 kb) under in vivo condition. Furthermore, we provide a summary of 44 genes currently investigated to demonstrate the general feasibility of this technique.Themes: Cellular and molecular biologyTopics: Gene structure and function: general Copyright (C) 1999 Elsevier Science B.V.
Original language | English |
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Pages (from-to) | 383-394 |
Number of pages | 12 |
Journal | Brain Research Protocols |
Volume | 4 |
Issue number | 3 |
DOIs | |
State | Published - Dec 1999 |
Bibliographical note
Funding Information:This work was supported by the Max-Planck-Society, the Human Frontier Science Program (RG 562/96) and the European Union (Bio4-98-0259) (SS). ON was supported by Axel Ullrich.
Keywords
- Alternative splicing
- Minigene
- RT-PCR
- Transfection
ASJC Scopus subject areas
- General Neuroscience