Identifying the Telomerase RNA of Aspergillus nidulans: the generation of a DNA construct and refinement of transformation procedures

Telomeres are genomic structures found at the ends of linear chromosomes that protect cells by ensuring the complete transmission of genetic information to future generations. As the cell replicates, the telomeres will naturally shorten, allowing them to be used as a biological clock. The enzyme telomerase, a complex molecule made of a protein (TERT) and an RNA component (TR), regulates telomere length. The TR region of the enzyme contains the template region used to synthesize the telomeric repeats, TTAGGG, which are found in most mammals as well as in our model organism of choice — Aspergillus nidulans. In this project, I attempt to knockout a putative TR template. Fusion PCR was used to synthesize a DNA construct that was transformed into fungal cells and integrated into the genome through homologous recombination. I hypothesize that this putative TR template mutation will result in an abnormal phenotype in the fungal cells that arises from the shortening of telomeres due to the dysfunctional telomerase.