UW Team Combines CRISPR With Duplex Sequencing to Detect Low-Frequency Mutations

SAN FRANCISCO (GenomeWeb) – Researchers from the University of Washington have developed a targeted sequencing method that couples the CRISPR/Cas9 system for target selection and a previously described method known as duplex sequencing, which improves the accuracy of next-generation sequencing.

The researchers are now using the method, which was described recently in a publication on the BioRxiv preprint server, in their translational oncology and forensics work and startup TwinStrand Biosciences, which spun out of UW earlier this year, has licensed it.

The CRISPR-duplex sequencing method builds off the duplex sequencing technique to analyze low-frequency mutations that the UW researchers first described in 2012. Duplex sequencing involves tagging both strands of DNA with random, but complementary, double-stranded primers. The molecules are then amplified, sequenced, and grouped by their tags. Errors that are introduced from PCR or sequencing can be filtered out by looking for discrepancies among molecules with the same tag. Two rounds of filtering are performed, one for each of the complementary tags.

The team has since used the method to show that it can identify very low-frequency mutations to the cancer gene TP53 in ovarian cancer patients.

However, the problem with duplex sequencing is that it is not very

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