dCas9 | Dissection of the mammalian transcription termination mechanism by CRISPRi technology.

Summary
RNA polymerase II (RNAPII) transcribes protein-coding genes in eukaryotes. The proper RNAPII transcription termination is crucial for generation of functional mRNAs, and then proteins. Termination, or stop of RNA synthesis, may occur before or after mRNA 3’-end cleavage and polyadenylation. In the last decade many details have been acquired about mRNA maturation, but still little is known about the termination per se. In this proposal I will redress this problem.
One of the proposed transcription termination triggers is RNAPII elongation complex pausing downstream of mRNA maturation signal. I will test this hypothesis, using sequence-specific protein binding in the living cells.
In the proposed project I will combine cutting-edge CRISPRi/dCas9 technology with classical molecular biology, high-throughput sequencing and computational biology. This approach will allow me to address my research objectives: (1) to elucidate the role of elongation complex pausing in transcription termination and (2) to characterise the induced pausing in terms of stress-resistance. Also by implementation of this project I will develop a bioengineering tool to induce elongation complex pausing.
This timely approach, taking advantage of innovative technologies, will allow me to elucidate previously unknown details of RNAPII transcription termination in mammals and expand our knowledge about this process. Importantly, my approach will allow studies on the chromosomal genes in the living cells, making the results unique and valuable.
Implementation of the proposed project will enhance my proficiency by improving and diversifying my skills in experimental and computational science as well as boosting my transferable skills, such as scientific communication and writing, project management and networking. This altogether will improve my competitiveness and potential on the way to independent career. Thus the proposal corresponds well with the aims of the Work Programme.
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More information & hyperlinks
Web resources: https://cordis.europa.eu/project/id/747391
Start date: 01-08-2017
End date: 31-07-2019
Total budget - Public funding: 183 454,80 Euro - 183 454,00 Euro
Cordis data

Original description

RNA polymerase II (RNAPII) transcribes protein-coding genes in eukaryotes. The proper RNAPII transcription termination is crucial for generation of functional mRNAs, and then proteins. Termination, or stop of RNA synthesis, may occur before or after mRNA 3’-end cleavage and polyadenylation. In the last decade many details have been acquired about mRNA maturation, but still little is known about the termination per se. In this proposal I will redress this problem.
One of the proposed transcription termination triggers is RNAPII elongation complex pausing downstream of mRNA maturation signal. I will test this hypothesis, using sequence-specific protein binding in the living cells.
In the proposed project I will combine cutting-edge CRISPRi/dCas9 technology with classical molecular biology, high-throughput sequencing and computational biology. This approach will allow me to address my research objectives: (1) to elucidate the role of elongation complex pausing in transcription termination and (2) to characterise the induced pausing in terms of stress-resistance. Also by implementation of this project I will develop a bioengineering tool to induce elongation complex pausing.
This timely approach, taking advantage of innovative technologies, will allow me to elucidate previously unknown details of RNAPII transcription termination in mammals and expand our knowledge about this process. Importantly, my approach will allow studies on the chromosomal genes in the living cells, making the results unique and valuable.
Implementation of the proposed project will enhance my proficiency by improving and diversifying my skills in experimental and computational science as well as boosting my transferable skills, such as scientific communication and writing, project management and networking. This altogether will improve my competitiveness and potential on the way to independent career. Thus the proposal corresponds well with the aims of the Work Programme.

Status

CLOSED

Call topic

MSCA-IF-2016

Update Date

28-04-2024
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Horizon 2020
H2020-EU.1. EXCELLENT SCIENCE
H2020-EU.1.3. EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions (MSCA)
H2020-EU.1.3.2. Nurturing excellence by means of cross-border and cross-sector mobility
H2020-MSCA-IF-2016
MSCA-IF-2016