Background
A gene that has been the focus of extensive pharmacogenomic research is the cytochrome P450 enzyme 2D6 (CYP2D6), which is highly polymorphic. Identifying how different alleles impact phenotype in terms of metabolism is desirable because CYP2D6 is involved in the metabolic pathway of up to 50 different drugs currently used in medicine. This project continues work that identified individuals with anomalous copy number calls1, that might indicate combinations of CYP2D6 with its adjacent gene (CYP2D7), known as hybrid alleles. Our objective is to identify precisely which such hybrid variants are present. In this manner, the improvement in technology gained will enable correct identification of a wider range of variants of this enzyme than was previously possible, for translation into clinical practice in the form of more accurate pharmacogenetics testing.
Methods
The methodology applied was a long-PCR approach to identify hybrid alleles of CYP2D6 using the technique described by Kramer et al. (2009)2 and Black et al. (2012)3, with fragment delineation by both agarose gel and Agilent 2100 Bioanalyzer (Agilent Technologies, Canada) electrophoresis.
Results
Results showed some successful amplification for the conditions established. Comparison made between different runs point to effective changes to the initial method. The technique employed is a modified version of that described by Kramer et al., using a buffer that is specific for GC-rich regions. Preliminary results have confirmed the method application in gene characterization, having successfully amplified structures inherent to *5-like genotype
Acknowledgements
BCH held an Undergraduate Research Initiative (URI) Summer Studentship and is now funded by an Alberta Centennial Addiction and Mental Health Research Chair fund held by KJA. A sample processed was collected as part of Workpackage 3 of the STOP study (www.stop-study.com), funded by the European Commission’s 7th Framework Programme, grant agreement 261411.