Hedgehog Signaling Protocols pp 67-78

Part of the Methods Inmolecular Biology™ book series (MIMB, volume 397)

Efficient Manipulation of Hedgehog/GLI Signaling Using Retroviral Expression Systems

  • Maria Kasper
  • Gerhard Regl
  • Thomas Eichberger
  • Anna-Maria Frischauf
  • Fritz Aberger

Abstract

Efficient manipulation of Hedgehog (HH)/GLI signaling activity is crucial to the analysis of molecular events underlying HH/GLI-regulated cell fate determination and tumor growth. In this article, we describe the use of retroviral expression systems as a valuable tool to activate or repress Hh-pathway activity in a broad spectrum of mammalian cells—including human cells—either by forced expression of the major Hedgehog-effectors GLI1 and GLI2 or by expression of the short-hairpin RNAs-targeting GLI mRNAs. We focus on two distinct retroviral systems that allow efficient and sustainable expression of GLI proteins in primary cells and cell lines of human origin: (i) a Moloney Murine Leukemia Virus-based and (ii) an HIV-derived lentivirus expression system, which allows transduction of both dividing and quiescent cells.

Key Words

Retroviral gene expression lentivirus Hedgehog signal transduction GLI proteins RNA interference 

Copyright information

© Humana Press Inc., Totowa, NJ 2007

Authors and Affiliations

  • Maria Kasper
    • 1
  • Gerhard Regl
    • 1
  • Thomas Eichberger
    • 1
  • Anna-Maria Frischauf
    • 1
  • Fritz Aberger
    • 1
  1. 1.Department of Molecular BiologyUniversity of SalzburgSalzburgAustria

Personalised recommendations