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Detect low-frequency HDR and NHEJ editing events

CRISPR-Quest Gene Editing Tests

The research community’s rapid acceptance of the CRISPR/Cas9 technology is propelling a stage of deep investment in technology development. Already, three companies have emerged focusing on CRISPR therapeutic applications: Intellia Therapeutics, Editas Medicine, and CRISPR Therapeutics. CRISPR-Quest Gene Editing Test is based on DiaCarta’s next-generation polymerase chain reaction technology – XNA technology. It provides a rapid, low-cost, ultra-sensitive quantification of both NHEJ- and HDR-editing events to meet the growing need for a screening technology that is generally applicable.

XNA technology has already been widely used for high-sensitivity and high-precision applications such as rare cancer mutation detection. Since HDR and NHEJ editing events can occur at very low frequency (<1%), especially HDR in primary or induced pluripotent stem (iPS) cells, XNA technology appears to be a fit for researchers wanting a rapid, sensitive, quantitative readout of editing in cells and tissues. The technique also enables empirical validation of guide RNA efficiency and measurement of the ratio of HDR: NHEJ at a targeted locus.

HIGHLIGHTS

  • Ultra-sensitive of low-frequency HDR and NHEJ editing events (powered by XNA technology)
  • Suitable for HDR in primary or induced pluripotent stem (iPS) cells
  • Enable empirical validation of guide RNA efficiency and measurement of HDR: NHEJ ratio at targeted locus
  • Rapid (assay run time < 2 hours)
  • Low cost (no capital investment like ddPCR and NGS)

Ordering information

Product NameCatalog # (RUO)Catalog # (CE/IVD)
CRISPR-Quest Gene-Editing Event Screening Kit (500 Reactions) DC-70-0500RN/A
CRISPR-Quest Gene-Editing Event Screening Kit (1000 Reactions)DC-70-1000RN/A

Note: RUO (Research Use Only) products are not for use in diagnostic procedures.

How it works

  • Nuclease-induced double-strand breaks (DSBs) can be repaired by one of the two pathways: Non-homologous end joining (NHEJ) and Homology-directed repair (HDR)
  • Imprecise NHEJ can produce insertion and/or deletion mutations of variable length at DSB site
  • HDR can introduce precise point mutations or insertions from ss- or ds- DNA donor template

Supporting data

qPCR Results
The rare cell with a single-base change is isolated using QClamp XNA-PCR as a measurement tool, then enriched sequentially until a pure clone resulted.

Calculated Percentage Modification
Compares favorably with amplicon sequencing and T7EI; Useful for small mutations and SNPs where T7EI may fail; Performs better on high % modification mixtures: this is useful for high-throughput modification workflow or clone screening.