I undertook an integrated 6.5 month full-time work placement in the Cytokine Signalling Unit at Institut Pasteur as part of my degree. During this work placement, I carried out two main research projects. Carrying out experiments for these projects allowed me to develop keen planning skills and learn advanced experimental design. The aim of the first of these projects was to identify and validate innate immune response targets of the interferon-induced microRNA, miR-3614-5p. I showed that miR-3614-5p silences the ADAR1 and STAT1 genes by binding to the 3’ untranslated region of their mRNA. Interestingly, miR-3614-5p appeared to positively regulate the innate immune gene CSF1R, which contrasts to the typical function of microRNAs.Through this research project, I gained competencies in many research techniques that include: mammalian cell culture, mammalian cell transfection, luciferase assay, PCR primer design, gel electrophoresis, DNA gel purification, cloning of DNA into plasmids, bacterial transformation and large-scale plasmid purification, site-directed mutagenesis, and DNA sequencing.The aim of the second research project was to utilise CRISPR-Cas9 genome editing to specifically edit the sequence of the rs12720270 single nucleotide polymorphism in the Janus Kinase TYK2 gene. This polymorphism is proposed to affect alternative splicing of the TYK2 transcript. Through this project, I gained many competencies, including: sgRNA design, CRISPR-Cas9 genome modification, isolation and selection of CRISPR-Cas9 clones, DNA extraction and purification, DNA sequencing.

I carried out a summer internship within the Corticosteroids Hormones in Cardiovascular Disease group, funded by a competitive Society for Endocrinology grant. The aim of this internship was to investigate variation within the 3’ untranslated region of the high blood pressure-associated MRAP gene in essential hypertensive patients. Variation within this region may affect microRNA binding and therefore MRAP gene expression. To investigate this variation, PCR primers for the MRAP 3’ untranslated region were designed and PCR using these primers and DNA from hypertensive patients was performed. Amplified DNA from hypertensive subjects were sequenced and variation within the MRAP 3’ untranslated region was identified using UGENE bioinformatic software. I found a single common polymorphism in this region of the MRAP gene of hypertensive subjects occurring at the same frequency as the general population. This polymorphism was not predicted to affect any microRNA binding sites. Through this research project, I gained many competencies, including: PCR primer design, PCR optimisation, gel electrophoresis, DNA sequencing.