title: The in vivo minigene approach to analyze tissue-specific splicing.
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.
authors.affiliation: Max-Planck Institute of Neurobiology, Am Klopferspitz 18a, D-82152, Martinsried, Germany.
authors.forename: O
authors.initials: O
authors.lastname: Stoss
authors.forename: P
authors.initials: P
authors.lastname: Stoilov
authors.forename: A M
authors.initials: AM
authors.lastname: Hartmann
authors.forename: O
authors.initials: O
authors.lastname: Nayler
authors.forename: S
authors.initials: S
authors.lastname: Stamm
mesh_terms: D017398:Alternative Splicing / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D005091:Exons; D005813:Genes, Synthetic / Q000235:genetics*; D005816:Genetic Complementation Test; D006801:Humans; D007438:Introns; D007668:Kidney / Q000166:cytology; D016296:Mutagenesis; D012322:RNA Precursors / Q000235:genetics; D014162:Transfection / Q000379:methods*
chemical_list: D012322:RNA Precursors