Understanding developmentally controlled co-transcriptional splicing in the mammalian nervous system
Award Number
BB/V006258/1Status / Stage
ActiveDates
1 July 2021 -30 June 2024
Duration (calculated)
02 years 11 monthsFunder(s)
BBSRC (UKRI)Funding Amount
£628,746.00Funder/Grant study page
BBSRC UKRIContracted Centre
King's College LondonPrincipal Investigator
Professor Eugene MakeyevPI Contact
eugene.makeyev@kcl.ac.ukPI ORCID
0000-0001-6034-6896WHO Catergories
Understanding Underlying DiseaseDisease Type
Dementia (Unspecified)CPEC Review Info
| Reference ID | 733 |
|---|---|
| Researcher | Reside Team |
| Published | 07/07/2023 |
Data
| Award Number | BB/V006258/1 |
|---|---|
| Status / Stage | Active |
| Start Date | 20210701 |
| End Date | 20240630 |
| Duration (calculated) | 02 years 11 months |
| Funder/Grant study page | BBSRC UKRI |
| Contracted Centre | King's College London |
| Funding Amount | £628,746.00 |
Abstract
Excision of introns from nascent pre-mRNAs has been proposed to control alternative splicing and the abundance of correctly processed transcripts. How developmental changes in co-transcriptional splicing patterns might contribute to rewiring of the gene expression program is an exciting open question. Building on our preliminary data we will test the hypothesis that the RNA-binding protein Ptbp1 promotes co-transcriptional excision of numerous introns and that its natural downregulation during neuronal differentiation facilitates a large-scale switch between co- and post-transcriptional splicing modes. We will also explore the possibility that this switch facilitates neuronal differentiation by altering expression of important target genes. We will pursue three interrelated objectives: (1) dissecting molecular mechanisms that allow Ptbp1 to activate co-transcriptional excision of introns; (2) elucidating the effect of co-transcriptional splicing on the abundance, isoform composition, and biological functions of Ptbp1 targets; and (3) understanding the role of Ptbp1 in co-transcriptional splicing dynamics in developing neurons. The first two objectives will involve in-depth analyses of Ptbp1-regulated candidates using in vitro and minigene-based assays, auxin-inducible depletion of Ptbp1, and differentiation of genetically modified cells into neurons. Our main model will be the Dnmt3b gene encoding a DNA methylase associated with cancer, immunodeficiency, developmental disorders and neurodegeneration. The third objective will combine unbiased sequencing approaches with viral vector-based Ptbp1 rescue experiments to address the extent to which this protein contributes to co-/post-transcriptional splicing transitions in developing neurons. Overall, this will uncover fundamental mechanisms linking pre-mRNA splicing and gene regulation in developing brain and delineate new possibilities for diagnosing and treating increasingly prevalent medical conditions.
Aims
We will pursue three interrelated objectives: (1) dissecting molecular mechanisms that allow Ptbp1 to activate co-transcriptional excision of introns; (2) elucidating the effect of co-transcriptional splicing on the abundance, isoform composition, and biological functions of Ptbp1 targets; and (3) understanding the role of Ptbp1 in co-transcriptional splicing dynamics in developing neurons.