Speakers
Description
Microphthalmia, Anophthalmia, and Coloboma (MAC) are severe ocular malformations resulting
from mutations in both protein-coding genes, such as the transcription factor SOX2, and non-coding
regulatory elements. Approximately 50% of affected individuals lack a definitive molecular diagnosis based
on exome sequencing aimed at detecting protein-coding genes mutations, suggesting that mutations in noncoding regions may play a critical role in disease pathogenesis.
We previously performed RNApolII-ChIA-PET analysis on neural stem cells derived from the mouse brain,
mapping long-range chromatin interactions between enhancers and gene promoters. These interactions
were subsequently mapped to the human genome, identifying 7.698 syntenic regions, termed human-mouse
syntenic long-range interactions (hmsLRI). DNA sequence variants associated with MAC (identified through
whole-genome sequencing of affected individuals) were then overlapped with this enhancer dataset.
This comparison identified significant copy number variants (CNVs) associated with genes involved in eye
development and inherited ocular diseases, though not affecting protein-coding regions; our study focuses
on four CNVs. We started from this selection to plan further functional studies, including transgenesis
experiments in zebrafish and the generation of brain and eye organoids from human pluripotent stem cells.
For in vivo analysis, CNVs were cloned into a Gateway vector, then transferred to a ZED vector optimized for
transgenesis in Danio rerio. This vector enables precise expression of transgenes and real-time monitoring
of gene activity using fluorescent reporters. These tools will provide valuable insights into how the identified
CNVs influence eye development and contribute to inherited ocular diseases
| Author(s) | Antoniazzi Gabriele1, Morciano Delia1, Marenco Carolina1, Pozzolini Giorgia1, Baldi Roberta1, Zakirova Elvira1, Mosconi Filippo1, D’Aurizio Romina2, Ceroni Fabiola3, Tabanera Noemi4, Beccari Leonardo4, Holt Richard3, Bovolenta Paola4, Ragge Nicky3 and Nicolis Silvia Kirsten1 |
|---|---|
| Affiliation(s) | "1 Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Milano, Italy 2 Institute of Informatics and Telematics (IIT), National Research Council (CNR), Pisa, Italy 3 School of Biological and Medical Sciences, Oxford Brookes University, Oxford, UK 4 Centro de Biología Molecular Severo Ochoa CSIC-UAM, Madrid, Spain" |
