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.
|Number of pages||12|
|Journal||Brain Research Protocols|
|State||Published - Dec 1999|
Bibliographical noteFunding 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.
- Alternative splicing
ASJC Scopus subject areas
- Neuroscience (all)