CRISPR and nanopore targeted sequencing for genotyping highly polymorphic genes.

Advances in sequencing and molecular genetic techniques have created an unprecedented opportunity for the development of personalized medicine. The impact of these advances has translated into improvements in diagnosis, prevention and, importantly, individualized therapeutic management. In the clinical context, genomic information from the individual has been routinely used in several areas, including transplantation medicine and pharmacogenetics. The implementation of new sequencing techniques in such contexts has the potential to address many of the limitations of conventional techniques, including level of resolution in highly polymorphic genes and sample processing time. With these considerations in mind, the overall objective of the present proposal is to evaluate the feasibility of typing highly polymorphic genes of clinical interest using nanopore sequencing and CRISPR-mediated enrichment. The long-term goal is to implement the use of next-generation sequencing technologies in the clinical context with a focus on precision medicine. The research question that the present study seeks to address is whether the results obtained by the nanopore-CRISPR methodology are comparable with conventional techniques for typing highly polymorphic alleles. The reasoning behind this question is that the implementation of such technologies would allow a more robust and accurate genotyping, in a shorter time and more affordable.

Medicina de precisión, medicina genómica, secuenciación por nanoporos, CRISP/Cas9, medicina de trasplantes, farmacogenómica, HLA, CYP450.