Crispr-Based Enrichment Strategies For Targeted Sequencing

The ability to easily produce or procure sequencing data has expanded to be within the reach of most clinics and research laboratories, but the complexity of sequence analysis remains a hurdle for many scientists, and a decline in sequencing cost means that the generation of gratuitous information in a given experiment is a challenge that is more and more often being encountered. To address this issue, methods have been present, some dating to the advent of nucleic acid sequencing, for capturing, targeting, or otherwise enriching specific nucleic acids in order to obtain greater depth of reads from a small portion of sequences within a complex sample. However, many of these methods have been complicated and laborious, relying on the design of hundreds to thousands of oligonucleotide probes, fabrication of microarray chips, and long hybridization times. Here, we review these methods, their benefits and uses, and catalog and discuss the implications of a recent development that has enabled a more efficient and expanded set of tools for enriching nucleic acids - the application of CRISPR technology. This introduction and analysis of the capabilities of new CRISPR-based enrichment strategies shows that it has the potential to expand the scope of enrichment to new possibilities, including the coupling of DNA and RNA targeting with long-read, portable sequencing platforms. Moreover, there are several areas where CRISPR-enrichment is a logical next step to more powerful and simplified sequencing for applications such as diagnostics and environmental monitoring.