Examining multiple cellular pathways at once using multiplex hextuple luciferase assaying.

ABSTRACT: Sensitive simultaneous assessment of multiple signaling pathways within the same cells requires orthogonal reporters that can assay over large dynamic ranges. Luciferases are such genetically encoded candidates due to their sensitivity, versatility, and cost-effectiveness. We expand luciferase multiplexing in post-lysis endpoint luciferase assays from two to six. Light emissions are distinguished by a combination of distinct substrates and emission spectra deconvolution. All six luciferase reporter units are stitched together into one plasmid facilitating delivery of all reporter units through a process we termed solotransfection, minimizing experimental errors. We engineer a multiplex hextuple luciferase assay to probe pathway fluxes through five transcriptional response elements against

SUBMITTER: Sarrion-Perdigones A 

PROVIDER: S-EPMC6911020 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature