Generation and characterization of a single-chain Fv antibody against G, a hedgehog signaling pathway transcription factor.
Gli3 is a key regulator of development, controlling multiple patterning steps. Here we report the generation of a scFv antibody specific to the repressor domain of human Gli3. We show that this scFv retains the binding capacity of its parent anti-Gli3 monoclonal antibody derived from hybridoma clone 5E1. When expressed in mammalian cells, the anti-Gli3 scFv co-localizes with intracellular Gli3. Immunocytochemical staining of the intrabody in Gli3-positive TM4 cells shows a distinct perinuclear cytoplasmic localization. Such a scFv constitutes a useful tool for studying transcriptional regulation of the hedgehog pathway in mammals and offers a starting point for developing novel Gli-related therapeutic intrabodies.
Laht
Silja
S
Celecure AS, Akadeemia tee, Tallinn, Estonia.
Meerits
Kati
K
Altroff
Harri
H
Faust
Helena
H
Tsaney
Robert
R
Kogerman
Priit
P
Järvekülg
Lilian
L
Paalme
Viiu
V
Valkna
Andres
A
Timmusk
Sirje
S
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000378:metabolism; D053823:Hedgehog Proteins / Q000378:metabolism*; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D007135:Immunoglobulin Variable Region / Q000737:chemistry*; D051741:Kruppel-Like Transcription Factors / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D000074290:Zinc Finger Protein Gli3
C465571:Gli3 protein, mouse; D053823:Hedgehog Proteins; D007135:Immunoglobulin Variable Region; D051741:Kruppel-Like Transcription Factors; D009419:Nerve Tissue Proteins; D014157:Transcription Factors; D000074290:Zinc Finger Protein Gli3