Stem cells and immunity

The workgroups has previously generated mutants in the Death Inducer Obliterator (DIDO) gene, and has shown a function for this gene in RNA transcription and splicing. DIDO gene mutants show global alterations in RNA splicing patterns, which we attribute to the absence of one of three isoforms (DIDO3). Recent data have shown that the levels of the DIDO3 isoform  control the production of differentially spliced RNA products in a large subset of genes (Mora Gallarod et al., 2019). Elimination of this isoform gives rise to splicing defects, leading to the production of a large complement of aberrant RNA molecules that give rise to an inflammatory response (Gutierrez et al., 2022). 

After supression of DIDO3 and activation of the response, we propose to target the downstream activated pathways with CRISPR. Objective is to generate mutants in which the alterations of RNA splicing remain present, but inflammatory signalling is supressed or interupted. In parralel, the nature of the abberant RNAs and downstream effectors such as NFkB, IRF3, and interferon-stimulated genes (ISGs) will be evaluated by complementary methods such as RT-PCR, western blotting and immunofluorescence. The relevance of the observations made during the practical work will be studied on the hand of public cancer databases (GEPIA2).  Finally, the student is expected to finish his/her project with a written report.