CRISPR-Cap: multiplexed double-stranded DNA enrichment based on the CRISPR system

Nucleic Acids Res. 2019 Jan 10;47(1):e1. doi: 10.1093/nar/gky820.

Abstract

Existing methods to enrich target regions of genomic DNA based on PCR, hybridization capture, or molecular inversion probes have various drawbacks, including long experiment times and low throughput and/or enrichment quality. We developed CRISPR-Cap, a simple and scalable CRISPR-based method to enrich target regions of dsDNA, requiring only two short experimental procedures that can be completed within two hours. We used CRISPR-Cap to enrich 10 target genes 355.7-fold on average from Escherichia coli genomic DNA with a maximum on-target ratio of 81% and high enrichment uniformity. We also used CRISPR-Cap to measure gene copy numbers and detect rare alleles with frequencies as low as 1%. Finally, we enriched coding sequence regions of 20 genes from the human genome. We envision that CRISPR-Cap can be used as an alternative to other widely used target-enrichment methods, which will broaden the scope of CRISPR applications to the field of target enrichment field.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • CRISPR-Cas Systems / genetics*
  • DNA / genetics*
  • Escherichia coli / genetics
  • Genome, Bacterial / genetics*
  • Genome, Human / genetics*
  • Humans
  • Sequence Analysis, DNA

Substances

  • DNA