in-Site

What is in-Site?

dGH in-Site™ Probes harness KromaTiD’s Directional Genomic Hybridization technology to provide whole genome tracking of inserted DNA cassettes as small as 2 kilobases. Unidirectional dGH probes provide a single cell, genome-wide perspective of cellular engineering outcomes, including tracking of viral and non-viral mediated insert integration (CRISPR/Cas and alternative systems).

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in-Site - All Channels: Normal
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Homolog 1 with bracketing probes (green) and insert (yellow) present

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Homolog 2 with only bracketing probes present (green)

Figure 1: Example of the dGH in-Site™ assay tracking on/off-target  integration in a CRISPR/Cas edited iPSC. Shown here in both homologs are two bracketing probes (green) on the centromeric and telomeric sides of the edit site. Insert probes (yellow) demonstrate on-target integration in one homolog (circled).

  • On- and off-target integration metrics
  • Clean integration data, even from complex or heterogeneous cell populations
  • A unique, whole genome, orthogonal method of direct visualization of inserts, without bioinformatic prediction of outcomes
  • Multi-channel fluorescence for flexible and multiplex assay design
  • Available for human, murine, canine, non-human primate and CHO cells.
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in-Site All Channels: Off-Target Integration
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Homolog 1 with bracketing probes (green) and insert (yellow) present

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Homolog 2 with only bracketing probes present (green)

Figure 2: Same dGH in-Site™ assay in a CRISPR/Cas edited iPSC, demonstrating both on-target and random integration of insert sequence (yellow) throughout the genome.

As an orthogonal analysis to PCR/sequencing techniques, dGH in-Site™ assays enable, through direct visualization, definitive measurement of the presence or absence of inserts. This includes genome-wide distribution and orientation of transgenes or inserted segments. When used in combination with our targeted probes and/or dGH dosimetry paints, dGH in-Site™ assays can provide quantitative, single cell on- vs off-target outcomes of the delivery and editing process, and can be used to track the stability of the edited genome in both clonal isolates and non-clonal populations.

Offering the lowest limit of detection of integrated or genomic DNA targets by fluorescence, dGH in-Site is the most comprehensive tool available for researchers interested in tracking engineering outcomes in a de novo fashion.

For more information on how dGH in-Site can inform your research, you can contact probes@kromatid.com or fill out the form below and we will reach out to you shortly!