Imaging alternative splicing in living cells.
We have developed an in vivo reporter of alternative splicing decisions that allows for the determination of FGF-R2 splicing patterns without the destruction of cells. This method has broad applications, including the study of other alternatively spliced genes in tissue culture and in whole animals, and may be useful in creating imaging markers for the study of tumor progression and metastasis. In this chapter, the authors present one example of this method using fluorescence reporters. As with any new assay, a series of experiments were performed to validate the method. This chapter documents some of these experiments.
Wagner
Eric J
EJ
Department of Biochemistry andBiophysics, University of North Carolina, Chapel Hill, USA.
Baines
Andrea
A
Albrecht
Todd
T
Brazas
Robert M
RM
Garcia-Blanco
Mariano A
MA
D017398:Alternative Splicing / Q000502:physiology*; D000818:Animals; D002478:Cells, Cultured; D005091:Exons; D049452:Green Fluorescent Proteins; D006801:Humans; D007438:Introns; D007668:Kidney / Q000378:metabolism*; D008164:Luminescent Proteins; D008297:Male; D008856:Microscopy, Fluorescence / Q000379:methods*; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism; D012322:RNA Precursors / Q000235:genetics; D051381:Rats; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism; D051497:Receptor, Fibroblast Growth Factor, Type 2; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics* / Q000378:metabolism
D008164:Luminescent Proteins; D012322:RNA Precursors; D017468:Receptors, Fibroblast Growth Factor; D049452:Green Fluorescent Proteins; C496353:FGFR2 protein, human; C496356:Fgfr2 protein, rat; D020794:Receptor Protein-Tyrosine Kinases; D051497:Receptor, Fibroblast Growth Factor, Type 2