Development of receptor and oncogene-responsive luciferase reporter vectors that are activated via kinase signalling to AP-1.

Authors

Danial J L Coleman, Luke Ames, Hurooul Aain, Joanna Gamble, Kinga Koscielniak, Zijin Zhang, Cherisse Lau, Abigail Joyce, Peter N Cockerill

Published in

Experimental hematology. Pages 105491. Aug 13, 2026. Epub Aug 13, 2026.

Abstract

The aim of this study was to generate lentiviral vectors carrying an array of AP-1 motifs driving luciferase gene expression as reporters of Mitogen Activated Protein Kinase (MAPK) activity. We created a series of vectors based on LeGO-iG that were used to generate stably transduced leukaemia cell lines. A vector termed LEGO-AP1 × 6-GM55 containing an array of 6 AP-1 sites linked to the minimal CSF2 promoter was sufficient to support high levels of MAPK-inducible luciferase activity in leukaemic cell lines that was suppressed by MAPK inhibitors. The inclusion of a putative chromatin priming element encompassing RUNX and ETS motifs increased the activity of these vectors. The additional inclusion of the full-length mouse CSF2 promoter, or the human DUSP5 promoter further increased the MAPK-dependent activity of these vectors in leukaemic cells. These vectors support moderate levels of constitutive activity in cells carrying mutations that activate the RAS/RAF/MEK MAPK signalling pathway, and high-level activity after direct activation of MAPK signalling. They also respond to T cell receptor activation via MAPK and Ca2+ signalling pathways. This resource will now make it easier to track receptor or oncogene-inducible MAPK activity in cultured cells, and potentially in tumours, in close to real time. TEASER ABSTRACT: Here we developed a series of lentiviral luciferase reporter vectors activated by RAS/RAF/MAPK signalling to AP-1. These vectors are activated in response to signalling driven by most of the signalling mutations detected in cancer cells or by receptors that control cell growth, differentiation and activation. These vectors have been optimised based on pathways known to activate gene expression in acute myeloid leukaemia or in activated T cells. The regulatory elements tested in these vectors includes an array of 6 AP-1 sites, the CSF2 promoter, the DUSP5 promoter and a chromatin priming element that binds RUNX and ETS factors.

PMID:
42595074
Bibliographic data and abstract were imported from PubMed on 14 Aug 2026.

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