IntroductionPersonalized Medicine and precision therapeutics hold promise to revolutionize modern clinical practice, by improving clinical decision making and maximizing drug efficacy, while minimizing drug toxicity, based on the unique patient’s genetic profile. Clinical decision support tools aim to help clinicians to implement genome-guided therapeutics and genomic medicine with the translation of a patient’s genomic information into a clinically meaningful format. Here, we developed a personalized genome interpretation workflow, based on open-source code for facilitating the practice of precision therapeutics.MethodsUsing two different previously validated pharmacogene panels, namely, the 12-pharmacogene PREPARE study panel and the 87-pharmacogene PyPGx panel, we comprehensively analyzed and reported clinically actionable, rare and novel variants, the majority of which lie at the introns and the fringe of the pharmacogenes in question.Results and DiscussionΤwo types of pharmacogenomics (PGx) reports were also produced for three members of a Greek family from data derived from whole genome sequencing. Our results demonstrate that this genome interpretation workflow enables targeted comprehensive clinical PGx assessment, with rare and novel PGx variants included in both reports for hypothesis generation. Although this overlap would not serve as validation, our workflow holds promise to facilitate implementation of PGx in the clinic using next-generation sequencing data.