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.
- 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)
|Product Name||Catalog # (RUO)||Catalog # (CE/IVD)
|CRISPR-Quest Gene-Editing Event Screening Kit (500 Reactions)||DC-70-0500R||N/A|
|CRISPR-Quest Gene-Editing Event Screening Kit (1000 Reactions)||DC-70-1000R||N/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
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.