Mechanism of transcriptional gene silencing induced by convergent transcription in human cells and its application in breast cancer.
Lead Research Organisation:
University of Oxford
Department Name: Sir William Dunn Sch of Pathology
Abstract
All organisms consist of cells that multiply through cell division. The genetic code, which defines each cell type, is stored within DNA molecules. DNA contains coding regions called genes. Genes are transcribed into messenger RNA (mRNA) molecules, which are in turn translated into proteins, a process called gene expression. It is important that each cell makes the right levels of proteins, which is achieved by controlled gene expression. Uncontrolled gene expression can lead to cancer or cell death. One of the most important mechanisms for the regulation of gene expression is gene silencing. Gene silencing in human cells requires the presence of small RNA molecules (siRNA), which activate the RNAi pathway. This leads to recognition and distruction of target mRNA in the cytoplasm. Another type of gene silencing is the establishment of specific modifications on DNA, which leads to inhibition of gene transcription, also called transcriptional gene silencing (TGS). Little is understood about the mechanism of TGS in humans. The work proposed in this fellowship application will use available knowledge about TGS from other organisms to investigate and explore the details of how TGS can be induced in humans and by what mechanism. This work has important implications given that it can lead to development of new research tools with a great potential for therapeutic application.
Technical Summary
All organisms consist of cells that multiply through cell division. The genetic code, which defines each cell type, is stored within DNA molecules. DNA contains coding regions called genes. Genes are transcribed into messenger RNA (mRNA) molecules, which are in turn translated into proteins, a process called gene expression. It is important that each cell makes the right levels of proteins, which is achieved by controlled gene expression. Uncontrolled gene expression can lead to cancer or cell death. One of the most important mechanisms for the regulation of gene expression is gene silencing. Gene silencing in human cells requires the presence of small RNA molecules (siRNA), which activate the RNAi pathway. This leads to recognition and distruction of target mRNA in the cytoplasm. Another type of gene silencing is the establishment of specific modifications on DNA, which leads to inhibition of gene transcription, also called transcriptional gene silencing (TGS). Little is understood about the mechanism of TGS in humans. The work proposed in this fellowship application will use available knowledge about TGS from other organisms to investigate and explore the details of how TGS can be induced in humans and by what mechanism. This work has important implications given that it can lead to development of new research tools with a great potential for therapeutic application.
Planned Impact
There is widespread interest in the transcriptional gene silencing (TGS) in human cells. However, very little is understood about the exact mechanism. The understanding of the properties of TGS in human cells is quickly outstripping the very limited amount that is known about induced gene expression regulation on transcriptional level. This project would therefore have an impact not only in biological fields, but also potentially in fields such as nanotechnology and biotechnology.
The project would open up fundamentally novel questions about TGS mechanism, and will therefore not only determine the role of TGS in gene regulation, but will also be relevant to development of a new research tool. This research proposal may be particularly interesting for scientific community, not only because of providing a new knowledge, but also for potential use of CT as a new tool for knocking down genes on transcriptional level in mammalian cells. There are no efficient commercially available tools for switching off the genes on transcriptional level at the moment.
The idea of CT induced TGS in human cells has been filed as patent by Isis Innovations at Oxford University and was received with great interest from various biotech companies. This project will help to develop a range of useful tools for scientific community as well as new therapeutic agents, which will be of great benefic for general public.
The project would open up fundamentally novel questions about TGS mechanism, and will therefore not only determine the role of TGS in gene regulation, but will also be relevant to development of a new research tool. This research proposal may be particularly interesting for scientific community, not only because of providing a new knowledge, but also for potential use of CT as a new tool for knocking down genes on transcriptional level in mammalian cells. There are no efficient commercially available tools for switching off the genes on transcriptional level at the moment.
The idea of CT induced TGS in human cells has been filed as patent by Isis Innovations at Oxford University and was received with great interest from various biotech companies. This project will help to develop a range of useful tools for scientific community as well as new therapeutic agents, which will be of great benefic for general public.
People |
ORCID iD |
Monika Gullerova (Principal Investigator / Fellow) |
Publications
White E
(2014)
Human nuclear Dicer restricts the deleterious accumulation of endogenous double-stranded RNA.
in Nature structural & molecular biology
Schlackow M
(2013)
Genome-wide analysis of poly(A) site selection in Schizosaccharomyces pombe.
in RNA (New York, N.Y.)
Schlackow M
(2013)
Understanding non-coding DNA regions in yeast.
in Biochemical Society transactions
Neve J
(2016)
Subcellular RNA profiling links splicing and nuclear DICER1 to alternative cleavage and polyadenylation.
in Genome research
Jin J
(2014)
p19-mediated enrichment and detection of siRNAs.
in Methods in molecular biology (Clifton, N.J.)
Burger K
(2015)
Swiss army knives: non-canonical functions of nuclear Drosha and Dicer.
in Nature reviews. Molecular cell biology
Burger K
(2017)
Nuclear phosphorylated Dicer processes double-stranded RNA in response to DNA damage.
in The Journal of cell biology
Burger K
(2018)
Nuclear re-localization of Dicer in primary mouse embryonic fibroblast nuclei following DNA damage.
in PLoS genetics
Burger K
(2019)
Tyrosine kinase c-Abl couples RNA polymerase II transcription to DNA double-strand breaks.
in Nucleic acids research
Bhardwaj S
(2016)
Transcription facilitates sister chromatid cohesion on chromosomal arms.
in Nucleic acids research
Description | Elucidating the mechanism of RNA dependent DNA damage response and its role in cancer |
Amount | £1,947,610 (GBP) |
Funding ID | 24866 |
Organisation | Cancer Research UK |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 01/2018 |
End | 12/2024 |
Description | Rosetrees Trust Seed award |
Amount | £10,000 (GBP) |
Organisation | Rosetrees Trust |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 01/2017 |
End | 12/2017 |
Description | Classification of endogenously produced small RNA |
Organisation | EMBL European Bioinformatics Institute (EMBL - EBI) |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | My lab covers experimental part of this project, supervision of the experiments as well as relevant consumables. |
Collaborator Contribution | Prof Enright provides bioinformatical expertise and supervision of Dr Davis. |
Impact | we generated data that are useful for other projects in my lab |
Start Year | 2014 |
Description | Dicer in alternative polyadenylation in humans |
Organisation | Manchester University NHS Foundation Trust |
Department | Biochemistry |
Country | United Kingdom |
Sector | Hospitals |
PI Contribution | Dr Kaspar Burger, PostDoc in my lab performed several experiments, which led us to create a model for Dicer function in alternative polyadenylation. We were meeting with our collaborators regularly to discuss obtained data and further development of the project. |
Collaborator Contribution | Dr Andre Furger's lab provided initial data set in this project, which included sequencing data of RNA isolated from nuclear and cytoplasmic fractions from wt and Dicer knockdown cells. |
Impact | Subcellular RNA profiling links splicing and nuclear DICER1 to alternative cleavage and polyadenylation. Neve J, Burger K, Li W, Hoque M, Patel R, Tian B, Gullerova M, Furger A. Genome Res. 2016 Jan;26(1):24-35. doi: 10.1101/gr.193995.115. Epub 2015 Nov 6. |
Start Year | 2014 |
Description | public lecture, TV and magazine interviews |
Form Of Engagement Activity | A magazine, newsletter or online publication |
Part Of Official Scheme? | No |
Type Of Presentation | Keynote/Invited Speaker |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | I have wide-ranging experience in disseminating information to a popular audience. As The Best Young Scientist of Slovak Republic (2008) and a winner of L'Oreal/UNESCO Women in Science Award UK and Ireland (2011) I have received significant media attention. I have extensive experience with radio and newspaper interviews in both countries, Slovakia and UK. Also, I gave a public lecture in Slovakia, which was broadcasted nation-wide. I received many emails from students and public, expressing motivation and interest in science in general and also my research. |
Year(s) Of Engagement Activity | 2009,2010,2011,2012,2013 |