Neovascular assays for the examination of antiangiogenic and antitumor effects of BAI1.
The transmembrane protein BAI1 is subject to extracellular proteolytic cleavage, rendering a 120kDa fragment that is capable of inhibiting angiogenesis. Emory University researchers have generated several plasmids and cell lines capable of expressing multiple forms of this gene, making them useful for studying the therapeutic potential of these proteins.
An uncleavable form of BAI1 is available that was generated through site directed alanine mutagenesis of the G-protein coupled receptor (GPS) proteolytic cleavage site (S927→A927). This uncleavable form of BAI1 has been cloned into the constitutively active pcDNA3 vector, where it has been sequenced for verification and stably transfected into HEK 293 cells. The 120kDa cleavage product of BAI1 is not detectable in conditioned media taken from transformants. Both the pcDNA3::BAI1A927 construct and HEK 293 cells transfected with this plasmid are available.
Plasmids and cell lines containing only the extracellular region of BAI1 are also available. The extracellular domain of BAI1 (a.a.1-929; truncated at GPS cleavage site) was cloned into pcDNA3 (pcDNA3::BAI1-ECD) and pTRE2 (pTRE2::BAI1-ECD) to drive constitutive and tetracycline driven expression, respectively. These vectors contain myc and His tags. Both pcDNA3::BAI1-ECD and pTRE2::BAI1-ECD are available in several different cell lines:
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