Investigating the biological roles of oxidised forms of 5-methylcytosine and active

由于在生物化学和遗传学的本科课程中学习了发育和发育障碍模块，我对从事科学研究产生了浓厚的兴趣，因此，在我的最后一年项目中，我研究了与细胞形状、极性和运动有关但机制尚不清楚的def 6基因（编码Rho GTPases的新型GEF）的结构与功能关系。应用分子生物学、细胞培养、转染和成像技术，我用COS-7细胞证明了DEF 6蛋白N端编码的PH样结构域对其正确的亚细胞定位和功能是必不可少的。虽然照顾生病的家庭成员影响了我的学位分类，但它增强了我追求生物医学研究事业的决心。因此，我申请并赢得了一个非常有竞争力的MRC资助的干细胞技术硕士研究生名额。通过这个极好的机会，我不仅获得了在所有三个胚层的衍生物中分离、培养、扩增和分化干细胞的高度相关的理论知识和实践技能，但更重要的是，我学会了如何制定一个有趣的生物学问题，采用正确的方法来解决它，同时认识到这种方法的局限性。通过运用这些技能，在我的硕士论文中，我能够首次成功地展示1- 5 mC氧化衍生物，5-hmC和5-caC在发育中的大脑中的空间分布和基因组定位，2-这些5 mC氧化衍生物瞬时积累，以表观遗传学重编程神经干细胞，使其经历神经到神经胶质的转换;这些5 mC氧化衍生物的瞬时积累在神经干细胞谱系特化过程中作为活性DNA去甲基化过程的中间体。从这些观察结果的定量分析到我们发现的报告，我了解到额外的高度相关的细胞报告。技能包括如何;实验和统计评估是否观察是显着的，审查文献，讨论我的意见，并成功地通过thechallenging审查过程，并提出他们给我的同行。我在我的书的第一章“贡献”中借鉴了这些技巧。

Thanks to the fascinating development and developmental disorders module during my undergraduatecourse in Biochemistry and Genetics, I became very interested in pursuing a career in scientific research.Accordingly, in my final year project, I studied the structure - function relationship of def6 gene (encodes anovel GEF for Rho GTPases) that was implicated in cell shape, polarity and movement but the mechanismwas unclear. By applying molecular biology, cell culture, transfection and imaging techniques I showed usingCOS-7 cells that the PH-like domain encoding N-terminal region of DEF6 protein is essential for its propersubcellular localisation and function. Although caring for an ill family member impacted my degreeclassification, it enhanced my resolve to pursue a career in biomedical research. As a result, I applied forand won a very competitive MRC funded MSc studentship in Stem Cell Technology. With this excellentopportunity, I have not only gained the highly relevant theoretical knowledge and practical skills ofisolating, culture, expansion, and the differentiation of stem cells in the derivatives of all three germ layers,but more importantly, I have learnt how to formulate an interesting biological question, employ the rightapproach to tackle it, while appreciating the limitation of such an approach. By employing these skills,during my MSc thesis, I was able to successfully show for the first time 1- The spatial distribution andgenomic localisation of 5mC oxidation derivatives,5-hmC and 5-caC in the developing brain, 2- These 5mCoxidation derivatives transiently accumulate to epigenetically reprogram neural stem cells undergoingneural to glial switch and 3- The transient accumulation of these 5mC oxidation derivatives serves asintermediates in an active DNA demethylation process during lineage specification of NSCs.From the quantitative analysis of these observations to the reporting of our findings Cell Reports I learnedadditional highly relevant skills including how to; experimentally and statistically evaluate whether anobservation is significant, review the literature, discuss my observations and successfully go through thechallenging review process and present them to my peers. I have drawn on these skills in my book chaptercontribution.