Search for microRNAs produced by gut bacteria with the potential to regulate host cell gene expression (360G-Wellcome-207183_Z_17_Z)

Through extensive analysis of the genome of gut commensal bacterium Bacteriodes spp., sequences predicted to form short (s)RNA hairpin structures homlogous to human microRNAs, and in particular miR-4787-5p. The target of this miRNA is ZBTB7A, a transcription factor involved in the transcriptional suppression of glycolysis. Prof Carding's group has previously shown that Bacteriodes spp. exports macromolecules including sRNAs in nanoscale microvesicles (outer membrane vesicles; OMV) that can be acquired by other gut bacteria and importantly for this project, intestinal epithelial cells. The aim of the project therefore is to determine if Bacteroides miR-4787-5p is structurally and functionally equivalent to mammalian miRNAs. The first goal of the project is to verify the sequence of the candidate miRNA, miR-4787-5p, through PCR and subsequent sequencing, and determine where in the bacterial cell it is expressed, and if it is present in OMVs. The next goal is to overexpress the miR-4787-5p sequences in a mammalian cell via a mammalian expression vector and determine the effect on expression of ZBTB7A, which is the target of the homologous mammalian miRNA, and the cells' metabolism and growth using moecular and cell based assays already established in the laboratory.