Cellular Engineering
Robust Structural Variation Detection & Genomic Integrity Assays
Careful Analysis of your Cell and Gene Therapy Research Approaches are Vital to an IND filing.
Chromosomal rearrangements are a source of structural variation within the genome that are prominent in human diseases, where there is extreme importance in detecting translocations and deletions. Traditional approaches have been used for detecting these inversions but are simply not robust enough to detect. Our dGH™ technology is a single stranded hybridization method, able to detect inversions routinely, providing high resolution using our chromosome paints.
As an orthogonal analysis to PCR/sequencing techniques, dGH 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 chromosome paints, dGH 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.
dGH in-Site™ Targeted Assays Provide Whole Genome Tracking of DNA Cassette Inserts as Small as 2kb.
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).
- On- and off-target integration metrics
- Detects structural variations in off target CRISPR therapies
- 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.
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.
dGH in-Site™ assay in an edited iPSC, demonstrating both on-target and random integration of insert sequence (yellow) throughout the genome.
Channel 1: Fluorescence channels overlaid, insert and bracketing probes visible on one copy of chromosome 22 and off target inserts in multiple chromosomes
Channel 2: Yellow fluorescence channel, insert probe signals on one copy of chromosome 22 (circled) and in multiple off-target chromosomes
Channel 3: Green fluorescence channel, bracketing probe signals on both copies of chromosome 22
dGH SCREEN™ Unbiased Assay Designed to Monitor Structural Variants throughout the Genome
With dGH SCREEN™, researchers are able to track and localize a wide variety of chromosomal rearrangements within a 10kb limit of detection. In addition to being an unbiased tool for the detection of structural variants throughout the genome SCREEN™ offers High Resolution Analysis of DNA repair activity by monitoring hallmarks of genomic instability, including sister chromatid changes.
Application Benefits of SCREEN™:
- Exchange Events & Orientation Events
- Chromosomal Gain & Loss Events
- Monitoring Cellular Engineering Outcomes
- Orthogonal Data for Sequencing
- Structural Integrity QC
- Variant Discovery
- Genomic Stability Assessment
This analysis produces vast amounts of rearrangement data which can be used to create stability and structural variance profiles for complex and heterogeneous cell populations, making dGH SCREEN one of the most comprehensive and robust karyographic assays available.
Example of the first step in the dGH SCREEN analysis process. A metaphase spread is hybridized with 5 color dGH probes designed to measure known and unknown variants in order to provide a whole genome survey of structural rearrangements in a single cell.
dGH SCREEN analysis example data. Chromosomes are organized karyographically and aggregate sample rearrangement data is gathered for analysis of genomic stability and structural integrity.
Develop a study plan for Genomic Integrity Karyotyping as a QC Method for your Cell Lines to Support your Cell Therapies
Analysis of 100+ samples for Genomic Integrity using G-Band Karyotyping for quality control assessment of your Investigational New Drug (IND).
- Develop a study plan with KromaTiD to use our assay for pivotal batch analysis.
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Comprehensive reports for each sample are prepared, summarizing the G-banding results and any identified chromosomal abnormalities.
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Genomic Integrity Reports are provided of a side-by-side comparison of your cell samples for your genomic integrity assessment and IND submission.