NEUROCRYSP · Regulation of cryptic splice sites in neuronal differentiation and disease
7РП — „Хора“ (Действия „Мария Кюри“)
- Период
- 2014-07-14 → 2016-07-13
- Финансиране от ЕС
- 231 283 €
- Участници
- 1
- Схема
- MC-IEF
Линиите свързват координатора с партньорите.
Накратко на български
Рекурсивното сплайсване в мозъка на гръбначните позволява премахването на много дълги участъци от генетичния код чрез междинни стъпки. Разбирането на този механизъм помага да се разбере как се регулира създаването на протеини при развитието на невроните и при различни заболявания.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Regulation of cryptic splice sites in neuronal differentiation and disease
NeuroCRYSP research project has studied splicing, in particular a complex splicing mechanism in vertebrates brain named Recursive Splicing. This mechanism allows splicing of long introns in the brain in a two-step model, as previously proposed for Drosophila. However, in vertebrates the process starts with the recognition of a cryptic RS-exon within the long intron. After splicing to its preceding intron, the RS-exon is skipped and both the upstream and downstream exons are joined together. The researcher in NeuroCRYSP project has contributed with many experimental approaches to understand recursive splicing mechanism in brain. These results were published in Nature journal in May 2015 and have greatly expanded knowledge about splicing mechanism in the scientific community. Moreover, in order to understand the function of recursive splicing further, the researcher has designed genome-editing experiments (experimental work performed by PhD rotation student Ms Andrea Elser) in order to disrupt recursive splice sites in induced pluripotent stem cells (iPSCs) and study their neuronal differentiation pattern. Recursive splicing generally ends up with RS-exon skipped from mature RNA. However, in some situations, such as within non-canonical minor isoforms expressed from alternative promoters, RS-exons can become included in mature RNA. Based on this observation, a new hypothesis was generated about the role of recursive splicing mechanism in the regulation of canonical splicing. The researcher performed many experimental and bioinformatics approaches to conclude that recursive splicing can affect splicing of many canonical exons in the transcriptome, which contain a 5’ splice site motif at the start of the exon. Moreover, the researcher found that recursive splicing inhibition by Exon Junction Complex (EJC) explains why canonical RS-exons normally remain included in mRNAs. We plan to submit these results soon to Cell journal. Mutations in genes coding different EJC components have been associated with rare diseases and neurodevelopmental disorders. The discovery of the involvement of EJC in regulation of recursive splicing is essential for future studies that focus on the molecular mechanism of these disorders and will potentially help to design therapeutic approaches to treat these diseases.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
A recent era of RNA research discovered complex RNA regulatory networks that involve RNA binding proteins (RBPs) and RNA. These networks are particularly dynamic and complex in the central nervous system, and can lead to neurologic diseases if deregulated. The host lab studies the regulatory networks that control alternative splicing in the brain. Recently, the lab identified thousands of cryptic splice sites that are bound by the spliceosome, but do not lead to active splicing in the adult brain. In the present project, I will assess if splicing at some of these cryptic sites is regulated during brain development or disease. Moreover, I will determine the importance of such regulation for neuronal differentiation.Most of the cryptic splice sites are present within long introns of genes that are only expressed in the brain. Therefore, I will employ genome-wide experimental and computational methods to study the regulation of cryptic splice sites in mouse embryonic stem cells (mESCs) and mouse brain from several developmental stages. It is known that binding of RBPs to target pre-mRNAs can actively repress cryptic splicing, which ensures expression of stable mRNAs. FUS and TDP-43 are two RBPs that regulate alternative splicing and lead to amyotrophic lateral sclerosis (ALS) when mutated. They have increased binding to the long introns and their depletion leads to decreased expression of long genes. I will therefore assess changes in splicing at cryptic sites upon depletion of FUS or TDP-43 in mESCs, in ALS mouse models, and in induced pluripotent stem cells (iPSCs) from ALS patients. Since de-repression of cryptic splice sites would lead to aberrant mRNAs, this may unravel the mechanism explaining how FUS and/or TDP-43 regulate the expression of long genes.The study of genome-wide cryptic splicing regulation might uncover a novel mechanism controlling neuronal development, and explain how misregulation of long genes contributes to ALS neuropathology.
Оригинален текст от CORDIS (на английски).
Участници
- UNIVERSITY COLLEGE LONDON · LONDONКоординаторнепознат регион
Връзки
Данни: CORDIS, © Европейски съюз