Efficient manipulation of Hedgehog/GLI signaling using retroviral expression systems.
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.
Kasper
Maria
M
Regl
Gerhard
G
Eichberger
Thomas
T
Frischauf
Anna-Maria
AM
Aberger
Fritz
F
D002460:Cell Line; D049109:Cell Proliferation; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D053823:Hedgehog Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D015513:Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D012190:Retroviridae / Q000235:genetics*; D015398:Signal Transduction; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic / Q000379:methods*; D014162:Transfection; D019076:Transgenes; D019065:Virus Assembly; D000071676:Zinc Finger Protein GLI1
D053823:Hedgehog Proteins; D015513:Oncogene Proteins; D015534:Trans-Activators; D000071676:Zinc Finger Protein GLI1