The Genomics Facility expertise and interest are focused in:

• Understanding the molecular bases of diseases, and namely the molecular events linking the NGF system (NGF and its receptors) to neurodegenerative diseases and chronic pain, along with the development of genomics, exploiting innovative biomarkers and pharmacological targets.
• Characterizing the pathways involved in the long-lasting analgesic effect of new therapeutic antibodies, such as anti-TRKA or anti-NGF in neuropathic pain.
• Transcriptomics analysis of neurodegenerative diseases biofluids (mainly Alzheimer’s, Parkinson’s diseases and Parkinsonisms) to identify biomarkers for an early and most accurate diagnosis and disease sub-type stratification.
• Cooperative studies involving an interdisciplinary approach, both laboratory and clinical setting for most accurate and early diagnosis towards a Personalized Medicine approach.

 

The skills acquired allow to contribute significantly to the realization and support of National, European and International project proposals.

The Genomics Facility offers innovative technologies and high-throughput genomic services through a varied portfolio of products and expertise:

 

Molecular biology

• DNA / RNA Extraction from cells, tissues, micro-vesicles;
• Qualitative and quantitative evaluation of nucleic acids (DeNovix, Bioanalyzer Agilent 2100)

 

NGS (Illumina)

• Library construction for next-generation sequencing (also low-input protocols);
• DNA-Seq, RNA-Seq, micro RNA Seq

 

Microarray on Agilent Technologies platform

• Profiling and analysis of differential expression of mRNA, microRNA and other non-coding RNA
• Comparative genomic hybridization (CGH array)

 

qRT Real Time- PCR (AriaMix, Agilent Technologies))

• Design and optimization of TaqMan and SybrGreen assays
• Differential Gene Expression Measurements and Analysis
• Expression of miRNA profiling
• Validation of target of interest obtained from RNASeq, miRNASeq experiments

 

Biomarker Measurements on cephalic-spine fluid (CSF) for neurodegenerative diseases, carried out by the Chemiluminescence method (CLEIA) with Lumipulse G600II (Fujirebio) device:

Beta-amyloid 1-42;

Beta-amyloid 1-40;

Total Tau;

p-tau

IL-6

 

Data analysis

The Facility applies and proposes standardized pre-analytical procedures in order to harmonize the results obtained, to identify validated international clinical criteria. The Facility strictly collaborates with the EBRI Bioinformatics facility (headed by Ivan Arisi) for data analyses and machine learning to detect patterns in large datasets to further enhance the capabilities of sequencing technologies.

 

Selected Publications

 

Pandolfini L, Barbieri I, Bannister AJ, Hendrick A, Andrews B, Webster N, Murat P, Mach P, Brandi R, Robson SC, Migliori V, Alendar A, D’Onofrio M, Balasubramanian S, Kouzarides T (2019) METTL1 Promotes let-7 MicroRNA Processing via m7G Methylation. Mol Cell. Apr 19. pii: S1097-2765(19)30269-2. doi: 10.1016/j.molcel.2019.03.040. Full Text

Terrigno M, Busti I, Alia C, Pietrasanta M, Arisi I, D’Onofrio M, Caleo M, Cremisi F. Neurons Generated by Mouse ESCs with Hippocampal or Cortical Identity Display Distinct Projection Patterns When Co-transplanted in the Adult Brain. Stem Cell Reports. 2018 Mar 13;10(3):1016-1029. doi: 10.1016/j.stemcr.2018.01.010. Epub 2018 Feb 15. Full Text

D’Onofrio M*, Arisi I*, Brandi R*, Malerba F, Paoletti F, Storti AE, Fulvio Florenzano, Luisa Fasulo and Cattaneo A (2014) proNGF/NGF mixtures behave as a new ligand in the NGF-regulated transcriptional response in PC12 cells”. BMC Neuroscience 2014 Apr 8;15:48. doi: 10.1186/1471-2202-15-48. Full Text