Lentiviral reporter constructs for fluorescence tracking of the temporospatial pattern of Smad3 signaling.
Canonical TGF-beta is involved in cell differentiation, tissue maintenance, and wound healing, but also plays a central role in the pathogenesis of diseases such as cancer Here we describe a lentivirus-based reporter vector system expressing green fluorescent protein (GFP) or red fluorescent protein (RFP) under the control of a Smad3-responsive element (CAGA)12 that allows observation of the temporospatial pattern of endogeneous Smad3-mediated signaling on a cellular level. Use of this method will be valuable to identify cells with active Smad3 signaling and investigate the role of endogenous Smad3 signaling in complex systems such as co-cultures in vitro, or in tumors during tumor cell invasion and metastasis in vivo.
Stuelten
Christina H
CH
Cell and Cancer Biology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-4254, USA. chrisstu@mail.nih.gov
Kamaraju
Anil K
AK
Wakefield
Lalage M
LM
Roberts
Anita B
AB
D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D011401:Promoter Regions, Genetic / Q000235:genetics; D015398:Signal Transduction; D051900:Smad3 Protein / Q000235:genetics / Q000378:metabolism*; D014162:Transfection / Q000379:methods
D051900:Smad3 Protein; D049452:Green Fluorescent Proteins