Stem cells and immunity
Centro Nacional de Biotecnología CSIC (CNB)
DESCRIPTION OF THE OFFER
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). While the role of the other two isoforms produced by the same gene (DIDO1 and DIDO2) still are unclear, recent data indicate that competition between DIDO2 and DIDO3 may control the production of differentially spliced RNA products in a large subset of genes (Mora Gallarod et al., 2019).
As DIDO gene organization makes genetic manipulation by classical methods unfeasible, we propose to target the DIDO2 isform with CRISPR. Objective is to generate DIDO2 mutants in which RNA splicing can be evaluated by classical means (RT-PCR) as well as by mini-genes containing a reporter enzyme. In parralel, the state of the RNA splicing machinery will be evaluated by complementary methods such as western blotting and immunofluorescence. Relevance of DIDO2 and DIDO3 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.
Biomolecules & Cell D.
Karel van Wely