Post-transcriptional events have been recognised as having a significant impact on the behaviour of the molecular machinery of the cells, inducing selective splicing, mRNA degradation in the cytoplasm and influencing cell fate determination, differentiation and proliferation. More specifically, the elevated activity of RNA editing and N6-methyladenosine (m6A), a methyl modification of adenosine, are ubiquitous in cancer cells.
Such post-transcriptional changes can occur anywhere in the transcriptome, however they are less often observed within open reading frames (ORF). When coding regions are edited, the aminoacid sequence and the expression of a protein are altered.
Post-transcriptional m6A is regulated through the activities of “writer”, “reader” and “eraser” genes. Writers catalyse the methylation; readers recognise the mehylated sites and decide on the fate of the modified mRNA, inducing, e.g., preferential binding to proteins, alternative splicing, post-translational modifications or protein degradation; erasers instead promote loci demethylation.
The advent of direct RNA sequencing, introduced by Oxford Nanopore Technologies (ONT), opens an avenue for the comprehensive, unbiased profiling of post-translational modifications.
The main goal of the Ph.D. is to investigate post-translational modifications cancer cell lines which so far have not been studied using ONT direct RNA sequencing and to functionally validate the findings using gene editing approaches such as CRISPR screens or interference.
Supervisor:
Dr. Giancarlo Russo
Contacts:
phone: +37052398249