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Interplay between DNA/RNA modifications and histone regulation
Sweden (IFMSA-Sweden) - Umea Universitet, Umea
Department of Medical Biosciences
Type of Research Project
- Basic science
What is the background of the project?
Epigenetics modifications are important for normal development and maintenance of tissue-specific patterns of gene expression. And the investigation of these modifications and modifiers is essential to uncover their implications during development and differentiation. LSD1 is a lysine-specific histone demethylase that removes mono and dimethyl group from H3K4 and H3K9. Several studies have indicated the importance of LSD1 in cell proliferation, adipogenesis, spermatogenesis, chromosome segregation and cancer. Furthermore, the contradictory roles for LSD1 in mouse embryonic stem cells (mESCs) have been also reported: some studies highlighted the importance of LSD1 in the maintenance of pluripotency whereas others in normal differentiation. To understand the role of the LSD1 in ESC and its interrelation with RNA and DNA modifications we have generated Lsd1 knockout mESCs using CRISPR/ Cas9 technology.
What is the aim of the project?
To generate Lsd1 KO mouse embryonic stem cells (mESC) To characterize LSD1 in mESCs. To study 6mA DNA and m6A RNA in Lsd1 KO mESCs.
What techniques and methods are used?
CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) / Cas9 Technology, cloning, cell culture, polymerase chain reaction, quantitative polymerase chain reaction, Western blot, Immunofluorescence.
What is the role of the student?
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The students will be shown how to perform each experiment in the beginning. Also, all the necessary guidelines will be given. Once the student learned the techniques required, he or she is expected to perform the experiments by herself or himself. They will also have to take care of cell lines (mESCs wild type and constitutive knock-out) required for the experiments. They are expected to participate in the lab meeting and discuss about the results. In addition, it is important for them to update their lab book daily.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
There are no mandatory courses, lectures. However, we will suggest students to read necessary articles related to the project and experiment. We will also explain theory behind the experiments.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a scientific report
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Previous experience with: molecular biology knowledge. Laboratory preferred.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts:
- Aguilo; F.; et al.; Coordination of m(6)A mRNA Methylation and Gene Transcription by ZFP217 Regulates Pluripotency and Reprogramming. Cell Stem Cell; 2015. 17(6): p. 689-704.
- Chen; T.; el at.; The lysine demethylase LSD1 (KDM1) is required for maintenance of global DNA methylation. Nature Genet; 2009. 41(1): p.125-9
- Wu; T.P.; Et Al.; DNA Methylation On N(6)-Adenine In Mammalian Embryonic Stem Cells. Nature; 2016. 532(7599): P. 329-33.
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