The in vivo minigene approach to analyze tissue-specific splicing

Oliver Stoss, Peter Stoilov, Annette M. Hartmann, Oliver Nayler, Stefan Stamm

Research output: Contribution to journalArticlepeer-review

51 Scopus citations

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 languageEnglish
Pages (from-to)383-394
Number of pages12
JournalBrain Research Protocols
Volume4
Issue number3
DOIs
StatePublished - 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

  • Neuroscience (all)

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