Cellular fibronectin toolset

Expression vectors encoding human 'cellular' fibronectin variants harboring the alternatively spliced Extra Domains B and/or A (or neither) ; fibronectin variant-expressing cells and purified recombinant cellular fibronectin variant proteins.

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Interest / Relevance: Cellular (also known as "oncofetal") fibronectin variants harboring one or both alternatively spliced so-called extra domains (EDB and EDA) play a central bioregulatory role during development, repair processes and fibrosis.

Keywords: extracellular matrix, fibrosis, angiogenesis, tumor microenvironment

Scientist's name: Ellen VAN OBBERGHEN-SCHILLING
Georgios Efthymiou
Agata Radwanska

Detailed technology overview: Detailed technology is described in :
Efthymiou G, et al. Fibronectin Extra Domains tune cellular responses and confer topographically distinct features to fibril networks. J Cell Sci. 2021 : jcs.252957 doi: 10.1242/jcs.252957 Published 1 February 2021.

- Construction of lentiviral expression vectors harboring the alternatively-spliced « cellular FN » variants containing Extra Domain A and/or B, or no Extra Domain (plasma-like FN)
- Production of lentiviral particles for transduction of FN variant-expressing cells
- Expression of FN variants in (assembly-incompetent) HEK293 cells
- Purification of soluble FN variants secreted by FN-expressing HEK293 cells
- Expression of FN variants in assembly-competent cells
- Addition of purified FN variants to cells for functional studies
- Preparation of variant-specific « cell-derived matrices »

Reference:

RT00508