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Fluorescence-based alternative splicing reporters for the study of epithelial plasticity in vivo.

Publication ,  Journal Article
Somarelli, JA; Schaeffer, D; Bosma, R; Bonano, VI; Sohn, JW; Kemeny, G; Ettyreddy, A; Garcia-Blanco, MA
Published in: RNA
January 2013

Alternative splicing generates a vast diversity of protein isoforms from a limited number of protein-coding genes, with many of the isoforms possessing unique, and even contrasting, functions. Fluorescence-based splicing reporters have the potential to facilitate studies of alternative splicing at the single-cell level and can provide valuable information on phenotypic transitions in almost real time. Fibroblast growth factor receptor 2 (FGFR2) pre-mRNA is alternatively spliced to form the epithelial-specific and mesenchymal-specific IIIb and IIIc isoforms, respectively, which are useful markers of epithelial-mesenchymal transitions (EMT). We have used our knowledge of FGFR2 splicing regulation to develop a fluorescence-based reporter system to visualize exon IIIc regulation in vitro and in vivo. Here we show the application of this reporter system to the study of EMT in vitro in cell culture and in vivo in transgenic mice harboring these splicing constructs. In explant studies, the reporters revealed that FGFR2 isoform switching is not required for keratinocyte migration during cutaneous wound closure. Our results demonstrate the value of the splicing reporters as tools to study phenotypic transitions and cell fates at single cell resolution. Moreover, our data suggest that keratinocytes migrate efficiently in the absence of a complete EMT.

Duke Scholars

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Published In

RNA

DOI

EISSN

1469-9001

Publication Date

January 2013

Volume

19

Issue

1

Start / End Page

116 / 127

Location

United States

Related Subject Headings

  • Wound Healing
  • Whole Body Imaging
  • Receptor, Fibroblast Growth Factor, Type 2
  • Protein Isoforms
  • Mice, Transgenic
  • Mice, Inbred C57BL
  • Mice
  • Keratinocytes
  • Fluorescence
  • Exons
 

Citation

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ICMJE
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Somarelli, J. A., Schaeffer, D., Bosma, R., Bonano, V. I., Sohn, J. W., Kemeny, G., … Garcia-Blanco, M. A. (2013). Fluorescence-based alternative splicing reporters for the study of epithelial plasticity in vivo. RNA, 19(1), 116–127. https://doi.org/10.1261/rna.035097.112
Somarelli, Jason A., Daneen Schaeffer, Reggie Bosma, Vivian I. Bonano, Jang Wook Sohn, Gabor Kemeny, Abhinav Ettyreddy, and Mariano A. Garcia-Blanco. “Fluorescence-based alternative splicing reporters for the study of epithelial plasticity in vivo.RNA 19, no. 1 (January 2013): 116–27. https://doi.org/10.1261/rna.035097.112.
Somarelli JA, Schaeffer D, Bosma R, Bonano VI, Sohn JW, Kemeny G, et al. Fluorescence-based alternative splicing reporters for the study of epithelial plasticity in vivo. RNA. 2013 Jan;19(1):116–27.
Somarelli, Jason A., et al. “Fluorescence-based alternative splicing reporters for the study of epithelial plasticity in vivo.RNA, vol. 19, no. 1, Jan. 2013, pp. 116–27. Pubmed, doi:10.1261/rna.035097.112.
Somarelli JA, Schaeffer D, Bosma R, Bonano VI, Sohn JW, Kemeny G, Ettyreddy A, Garcia-Blanco MA. Fluorescence-based alternative splicing reporters for the study of epithelial plasticity in vivo. RNA. 2013 Jan;19(1):116–127.

Published In

RNA

DOI

EISSN

1469-9001

Publication Date

January 2013

Volume

19

Issue

1

Start / End Page

116 / 127

Location

United States

Related Subject Headings

  • Wound Healing
  • Whole Body Imaging
  • Receptor, Fibroblast Growth Factor, Type 2
  • Protein Isoforms
  • Mice, Transgenic
  • Mice, Inbred C57BL
  • Mice
  • Keratinocytes
  • Fluorescence
  • Exons