NEURORNATRANSPORT · Molecular mechanims of mRNA transport in neurons
7РП — „Хора“ (Действия „Мария Кюри“)
- Период
- 2012-07-01 → 2014-06-30
- Финансиране от ЕС
- 200 372 €
- Участници
- 1
- Схема
- MC-IEF
Линиите свързват координатора с партньорите.
Накратко на български
Молекулярните механизми на транспорт на мРНК в невроните се изучават чрез модел с плодови мухи. Това помага да се разбере как клетъчното разпределение на генетичната информация контролира процеси като миграцията на клетките и предаването на сигнали.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Molecular mechanims of mRNA transport in neurons
Transport of mRNAs into defined subcellular compartments coupled to their local translation is a powerful mechanism to control gene expression in time and space. Originally believed to be a rare process involving only a handful of transcripts in highly specialised cells, more recent evidence suggests that potentially hundreds of transcripts are localised to specific subcellular compartments in a wide range of cell types. Moreover, mRNA localisation has been shown to be functionally important for diverse cell biological processes, such as cell polarity, cell migration, asymmetric cell division and signal transduction. However, the molecular mechanism governing mRNA localisation is currently unclear. In this project we aim to shed light on the regulation and functional significance of mRNA transport using the genetically tractable organism Drosophila melanogaster as a model system. We have developed a sensitive protocol that allows detection of mRNPs in the intact nervous system of the Drosophila larva. We have used this assay to conduct a screen in fly sensory neurons of 300 genomic 3’UTRs for dendritic or axonal localisation signals. We found that although low level dendritic localisation is widespread, efficient localisation into dendrites is a rare process. We identified a single 3’UTR that harbours a strong dendritic localisation signal. We have mapped a region within this 3’UTR that is absolutely required for dendritic localisation and now use this information to test for the functional significance of the process by genome engineering. To this end we have developed a powerful toolbox for CRISPR/Cas genome engineering in Drosophila. The toolbox consists of transgenic Cas9 fly strains and high efficiency gRNA vectors. In a proof-of-principle study we have demonstrated that these reagents allow genome engineering with remarkable high efficiency, including precise modifications of the genome by homology-directed repair. We now apply this technology to study in detail dendritic mRNA localisation of our candidate gene
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
RNA transport coupled to local translation presents an efficient means to regulate gene expression in time and space. Recent studies have indicated that this mode of post-transcriptional regulation is used in many different cell types and applied to hundreds to thousands of transcripts. mRNA transport is of particular importance in neurons, where it is involved in processes such as axon guidance and activity-induced synaptic plasticity. Loss of proteins involved in regulating RNA transport have been linked to hereditary mental retardation and dendritic targeting of certain mRNAs has been shown to be crucial for formation of long-term memories. However, which mRNAs are differentially localized in neurons and by what mechanism they are transported remains largely unknown.Here we propose to investigate this problem using the model organism Drosophila melanogaster, which offers significant experimental advantages, such as little genetic redundancy, powerful tools for loss- and gain-of-function studies and the possibility of visualize living neurons in their natural environment at high resolution. Furthermore, previous work has established that the overall mechanism of RNA transport is conserved from fly to humans. Thus, findings in the fly system are likely to have direct implications for our understanding of the human brain.We will adapt an established live cell imaging approach to identify mRNAs that are specifically enriched in neuronal processes. Identified transcripts will be characterized in further detail and used to search for RNA-binding proteins that specifically contact their localization elements. Candidate proteins will be tested for a role in mRNA transport using genetics coupled to high resolution imaging. Eventually, proteins regulating mRNA localization will be tested for a physiological role in the brain. Our work should reveal new insights into the cell biology of neurons and have broad implications for neuronal development, plasticity and disease.
Оригинален текст от CORDIS (на английски).
Участници
- MEDICAL RESEARCH COUNCIL · LONDONКоординаторОбединеното кралство
Връзки
Данни: CORDIS, © Европейски съюз