Offerte TESI PER I CORSI DI LAUREA MAGISTRALE

NF-Y is a trimeric Transcription Factor involved in activation of many "cancer" genes, by binding to the CCAAT box of their promoters. We constructed  ...

NF-Y is a trimeric Transcription Factor involved in activation of many "cancer" genes, by binding to the CCAAT box of their promoters. We constructed a comprehensive database of epithelial tumors (including but not limited to TCGA) with clinical, genetic and epigenetic data. We are currently systematically interrogating it for expression of the three subunits, as well as the NF-Y regulome (targeted genes) in cancer vs normal samples.

The student will deal with these vast interrogations, as well as with NF-Y expression and regulome in haematological tumors, for whichlittle is known.

We study the function of the NF-Y transcription factor in mouse Embryonic stem cells (mESCs). Sepcifically, we are interested in the splicing isoforms ...

We study the function of the NF-Y transcription factor in mouse Embryonic stem cells (mESCs). Sepcifically, we are interested in the splicing isoforms (NF-YAl and NF-YAs) of the regulatory subunit NF-YA. We have edited one exon in mESCs and observe the lack of mesoderm differentiation in the edited clones, as evidenced from RNA-seq and single cell analysis.

This indicates that NF-YAl is required for mesoderm specification and the regulome of NF-YAl in these cells needs now to be understood.

The student will charcaterize this phenomenon at the molecular and cellular level with common molecular biology techniques (RT-PCR, ChIP, Western blot, IFs, FACS).

NF-Y is a trimeric transcription factor present -and extremely conserved in all eukaryotic. The three genes (NF-YA/B/C) have been enormously expanded  ...

NF-Y is a trimeric transcription factor present -and extremely conserved in all eukaryotic. The three genes (NF-YA/B/C) have been enormously expanded in plants. The systematic organization of NF-Y genes were first done by us in Arabidopsis thaliana. Studies in other plants (rice, maize, canola etch) have since followed. Dozens of genetic experiments -overexpressions and inactivations of single subunits in various plant species- have been performed.

The sequence of several hundreds plants genomes are now available: the student will reconstruct the phylogenetic history of the NF-Y genes in  plants by comparative genomics approaches and relate the data to the available genotype/phenotype experiments in vivo.

Regulation of gene expression is governed by transcription factors (TFs) which bind to discrete genomic locations. Many protooncogenes and tumor suppr ...

Regulation of gene expression is governed by transcription factors (TFs) which bind to discrete genomic locations. Many protooncogenes and tumor suppressors are TFs and their dysregulation leads to altered gene expression resulting in uncontrolled cell growth and cancer.

The CCAAT box is a common DNA element, critical for the expression of many cell-cycle and cancer related genes. It is specifically bound by NF-Y, a pivotal transcriptional regulator known to synergise with and to facilitate TF binding to DNA acting as a “pioneer” factor. Genome wide analyses of TF bound regions have shown that NF-Y is frequently associated with a discrete set of TFs, among which the bHLH, bZIP and E2F families of protooncogenes Myc, Fos and E2F1.

The proposed thesis research topic aims at understanding the molecular interactions of NF-Y with other TFs, at the biochemical, structural and functional levels, based on “prototypical” promoters bound by NF-Y and its genomic partners, which are relevant in cancer progression.

Techniques:

Molecular Biology: cloning, generation of recombinant fusion protein expression vectors

Molecular Genetics: EMSA, mammalian cell colture, transfections, Luc reporter assays; ChIP; qRT-PCR; ImmunoPrecipitation, Western Blotting

Biochemistry: recombinant protein purification for structural and biophysical analyses (crystallography, CryoEM, SAXS, DLS, RPI)