WP 6.1
Technological approaches
for RNA therapeutics

General objectives

This Workpackage aims at providing a multimodal and multistage approach to RNA therapeutics by sequencing, imaging and editing of RNA coupled with tailored computational and artificial intelligence approaches. The aim is to decipher the complexity of human transcriptome and epitranscriptome by means of technologically advanced approaches.

Methodology

Molecular and cell biology methods for RNA editing: Nanopore direct RNA Sequencing and Artificial intelligence analysis, confocal, image scanning, multiphoton, super resolved and label-free CIDS; Image scanning microscopy, fluorescence fluctuation spectroscopy, fuorescence lifetime assay to understand the chemical environment of the RNA therapeutic.

Task 6.1.1

Task 6.1.2

Task 6.1.3

Task 6.1.1

Task Leader: Graziano Pesole
PI: Graziano Pesole, Silvestro Conticello, Ernesto Picardi, Valter Tucci, Barbara Storti

PI: Barbara Storti

Validate the most appropriate RNA editing strategy for inducing synthetic lethality in tumor cells in specific transcript target (induction of stop codon, alteration of splicing, etc.) in cellular and animal models by imaging (high-resolution microscopy, nanoscopy) on established cellular models of cancer. Relevant Publications (2022-2023) Nepita, I.; Piazza, S.; Ruglioni,...

PI: Valter Tucci

In recent years we witnessed a fast revolution in gene-editing technologies, such as clustered regularly interspaced palindromic repeats (CRISPRs) and CRISPR-associated (Cas) proteins. CRISPR technology is based on DNA breaks and homology-directed repair mechanisms, which can be programmed by noncoding single guide RNAs (sgRNAs) to guide specific nuclease (e.g., Cas9...

PI: Ernesto Picardi

RNA editing is a relevant epitranscriptome mechanism by which cellular RNAs are modified in specific localizations in a not-transient fashion. In humans, RNA editing involves the deamination of cytosine in uridine by APOBEC enzymes and, most frequently, the conversion of adenosine in inosine by the ADAR family of enzymes (PMID:...

PI: Silvestro Conticello

Objective: Development of programmable RNA editing as a tool to interfere with specific cellular processes and trigger selective cell death in cancer by targeting cellular processes in a synthetically lethal relationship with processes already altered in the cancer cells. Synthetic lethality is a condition where simultaneous occurrence of two specific...

PI: Graziano Pesole

RNA editing is a relevant epitranscriptome mechanism by which cellular RNAs are modified in specific localizations in a not-transient fashion. In humans, RNA editing involves the deamination of cytosine in uridine by APOBEC enzymes and, most frequently, the conversion of adenosine in inosine by the ADAR family of enzymes (PMID:...

Task 6.1.2

Task Leader & PI: Francesco Nicassio

PI: Francesco Nicassio

We aim at exploiting Oxford Nanopore direct RNA Sequencing (DRS) to dissect the complexity of human cancer transcriptome and epitranscriptome. We will adopts cutting edge experimental and computational approaches to provide key insights into the complexity, modification profile and regulation of cellular RNAs, with a particular focus on non-coding RNAs...

Task 6.1.3

Task Leader: Alberto Diaspro
PI: Alberto Diaspro, Giuseppe Vicidomini

PI: Giuseppe Vicidomini

Development in progress

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PI: Alberto Diaspro

Development in progress

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