The in vivo minigene approach to analyze tissue-specific splicing

  • Stoss O
  • Stoilov P
  • Hartmann A
 et al. 
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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.

Author-supplied keywords

  • Alternative splicing
  • Minigene
  • RT-PCR
  • Transfection

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  • Oliver Stoss

  • Peter Stoilov

  • Annette M. Hartmann

  • Oliver Nayler

  • Stefan Stamm

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