John Pooley

Background

About John Pooley

Five years research experience in gene therapy using adeno-associated virus (AAV) vectors and CRISPR-Cas9. Versatile, adaptable, capable of learning new areas at pace, setting up new laboratory methods from scratch, and highly advanced benchwork troubleshooting ability. Setup & run AAV manufacture, endotoxin testing, digital droplet PCR and explant culture models. Extensive experience with quantitative PCR and RT-PCR, western blot, immortalised cell & primary cell culture, plasmid cloning & production (restriction enzymes, inverse PCR, Gibson assembly, golden gate, site directed mutagenesis), transduction and transfection of plasmids and AAV into cells and tissues, chromatin immunoprecipitation, confocal and widefield microscopy (including advanced methods FRET & number and brightness assay), fluorescent immunohistochemistry, flow cytometry, co-immunoprecipitation, detailed statistical knowledge & execution, awareness and application of some next-generation sequencing technologies and analyses. My current work has led to the development of novel gene therapy strategies for glaucoma and post-haemorrhagic hydrocephalus, and furthered development of an existing CRISPR-Cas9 technology for these conditions. For several years I have worked with an industry partner to perform directed evolution of AAV capsids and novel promoter development for specific targeting of these candidate therapeutics in eyes. I have awareness of prior art and the intellectual property space, can compose successful large research grant applications, and have developed proposals involving phase I trial models. A further 12 years research experience in glucocorticoid (stress hormone) biology and rhythms, including chromatin biology. My work here helped to understand the role of the hourly rhythm of glucocorticoid secretion in mediating fine control of gene expression and define stress reactive binding sites for glucocorticoid receptors in brain tissue. I further redefined interactions between glucocorticoid receptors and similar proteins and how they shape gene regulation. Having built a research portfolio, substantial experience and adaptability I would now like to explore research and non-research opportunities outside of the academic/University setting. In addition to highly developed laboratory skills my attributes also include; project management, scientific writing, supervision/teaching & staff management, oral & written presentation skills, budget management, communication of science to lay and expert audiences, strong organisation skills, problem solving and idea generation.