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 an 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 an 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 and we will reach out to you shortly!