MarkerGene™ Cell Proximity Assay Kit
Cell-cell communication is an essential facet of growth and development for higher organisms. These interactions are responsible for a multitude of biological regulatory functions including cell-cell contact inhibition, organ development, and cell movement. Cell-cell interactions also play a role in numerous pathologies such as metastasis initiation and host-pathogen infection mechanisms. This kit contains reagents and methods to measure cell proximity using two popular marker genes, beta-galactosidase (lacZ) and our patented red luciferase (rluc). The lacZ gene is cloned into one cell line and rluc gene into the other then both cell lines are cultured together. Following treatment of the cells with our proprietary cell proximity substrate, cell contact and their corresponding interactions can be monitored.
When cells come into close proximity the lacZ+ cell releases a chemiluminescent substrate that can diffuse into neighboring rluc+ cells. The resulting red light emission (λ = 619 nm) can be used to give a quantitative measure of cell proximity. We have demonstrated the principle of the technique in a variety of cells grown in culture. Only when both cells are mixed is light emission exhibited in a cell number dependent manner.
- Sellmyer M, Bronsart L, Imoto H, Contag C, Wandless T, Prescher J (2013) "Visualizing cellular interactions with a generalized proximity reporter" Proc Natl Acad Sci 110(21):8567"?8572.>
- Jones KA, David J. Li DJ, Hui E, Sellmyer MA, Prescher JA, (2015) "Visualizing cell proximity with genetically encoded bioluminescent reporters" ACS Chem. Biol. , 10: 933-938.
- Porterfield WB, Prescher JA (2015) "Tools for visualizing cell-cell interactomes." Curr. Opin. Chem. Biol. 24: 121-130.
- Germain RN, Robey EA, Cahalan MD (2012) "A decade of imaging cellular motility and interaction dynamics in the immune system." Science 336: 1676-1681.
- Paley MA, Prescher JA (2014) "Bioluminescence: A versatile technique for imaging cellular and molecular features." Med. Chem. Comm. 5: 255-267.
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