High Throughput Screening

Overview

The High-Throughput Screening (HTS) Facility was established at the ICGEB Trieste in 2008 as a result of a collaborative project involving four research centres in Northeast Italy (ICGEB Trieste, LNCIB, SISSA, CIME), financed by the Friuli-Venezia-Giulia Regional Government.

The main goal of the High-Throughput Screening (HTS) Facility is to provide the research community with a platform of up-to-date instrumentation and adequate expertise to apply high-throughput screening approaches, in particular genome-wide RNA interference screening (siRNA miRNA) and drug repurposing screening, to fundamental biomedical research. The HTS Facility at the ICGEB has been operative since 2008 and its research activity has been horizontal to multiple success scientific stories within ICGEB network.

Research Output:

Buratti, E. et al. Deferoxamine mesylate improves splicing and GAA activity of the common c.-32-13T>G allele in late-onset PD patient fibroblasts. Mol Ther Methods Clin Dev 20, 227-236, doi:10.1016/j.omtm.2020.11.011 (2021).

Papa, G. et al. CRISPR-Csy4-Mediated Editing of Rotavirus Double-Stranded RNA Genome. Cell Rep 32, 108205, doi:10.1016/j.celrep.2020.108205 (2020).

Moimas, S. et al. miR-200 family members reduce senescence and restore idiopathic pulmonary fibrosis type II alveolar epithelial cell transdifferentiation. ERJ Open Res 5, doi:10.1183/23120541.00138-2019 (2019).

Rehman, M. et al. High-throughput screening discovers antifibrotic properties of haloperidol by hindering myofibroblast activation. JCI Insight 4, doi:10.1172/jci.insight.123987 (2019).

Gabisonia, K. et al. MicroRNA therapy stimulates uncontrolled cardiac repair after myocardial infarction in pigs. Nature 569, 418-422, doi:10.1038/s41586-019-1191-6 (2019).

Ali, H. et al. Cellular TRIM33 restrains HIV-1 infection by targeting viral integrase for proteasomal degradation. Nat Commun 10, 926, doi:10.1038/s41467-019-08810-0 (2019).

Genome-wide RNAi screening identifies host restriction factors critical for in vivo AAV transduction. Proc Natl Acad Sci USA, 112(36):11276-81.  doi: 10.1073/pnas.1503607112 (2015).

Instruments:

• PerkinElmer Operetta CLS (8 LED, Confocal, Enviromental Control (CO2, temperature), LWD objectives and High NA water immersion objectives ranging from 5x to 63x);
• PerkinElmer Operetta (Confocal, Enviromental Control (CO2, temperature), Long working distance objectives, High Numerical Aperture objectives ranging from 2x to 40x);
• PerkinElmer Envision multimode plate reader with dual monochromator, dual injector and temperature control;
• GX Collaborative Robotic Arm, PAA;
• Hamilton Starlet, 96 channel head, 2 independent channels;
• Plate washer Biotek 405;
• 2 x Multidrop multimode dispenser THermoFisher;
• Dedicated cell culture room (BSL2)
• Thermo Fisher ALPS Manual microplate sealer.

Arrayed Libraries:

• Mouse whole genome siRNA library (19,059 SmartPool siRNAs, Dharmacon);
• Human whole genome siRNA library (18,120 SmartPool siRNAs, Dharmacon);
• Human miRIDIAN microRNA mimics (Dharmacon) miRbase release V19. (2042 molecules);
• 1200 FDA approved small molecules (Prestwick Chemical);
• 320 Phytochemical small molecules (Prestwick Chemical);
• 480 Smart Fragment of FDA approved drugs (Prestwick Chemical);

Data Analysis:

• Harmony image analysis software, PerkinElmer
• Columbus server version of Harmony software, PerkinElmer
• Ingenuity Pathway Analysis Software, Qiagen