dGH in-Site™

Examine Edits One Cell at a Time

dGH in-Site™ gives researchers the ability to directly visualize and characterize rearrangements at their edit sites. Using synthetic oligo probes and repeat-free bioinformatic design, dGH in-Site is capable of tracking edit site translocations, inversions, copy number variants, viral integrations and other complex rearrangements with an ≈2kb resolution.

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 chromosome 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.

Concerns over a safe edit site? dGH in-Site™ allows researchers to examine their edits one cell at a time without having to worry about PCR biases or misrepresentative sequencing averages from heterogeneous cell populations. An orthogonal and complementary technique dGH in-Site™ to PCR/sequencing techniques helps provide the most comprehensive data available.

1/4 -Example of in-Site and SCREEN combination assay in unedited human fibroblast control line cell. Bracketing in-Site probes on chromosome 22 (circled) to identify insert target location and dGH SCREEN paints on chromosomes 1, 2, and 3 (pink). No insert would be expected in this control line
2/4 - Example of a single cell from the same in-Site assay in an edited iPSC viewed in multiple fluorescence channels. Left image shows all fluorescence channels overlaid, middle shows yellow/insert probe channel, and right shows green/on-target bracketing probe channel. Multiple inserts are visible outside of the on-target bracketing probes, indicating high levels of off-target integrations in this cell.
3/4 - Example of the same in-Site assay in an edited iPSC line cell with no off target insertions. Target site bracketing probes in green and insert probe in yellow zoomed in and expanded on the right. Results from this sample indicated that 6% of cells had no off-target insertions.
4/4 - Image from a dGH in-Site assay in polyploid, dual-edited, HEK-293FT cells demonstrating both normal and rearranged edit site structures. At the normal edit sites (A), both sets of in-Site probes are located on the same side of the chromosome. An inversion at the edit site is also visible (B), with one portion of the in-Site probe switching orientations and showing up on the opposite chromatid from the other portions of the in-Site probe. A partial deletion at one edit site is also observed (C), with the red portion of the in-Site probes missing.

Ordering Information

CATALOG	PRODUCTS	LIST PRICE
DGH-001	in-Site Adherent/Suspension Culture Development: Thaw, recovery, and harvest optimization	$1,000.00
DGH-002	in-Site Standard Probe Production: Design and verification of standard probes	$1,100.00
DGH-003	in-Site Assay Execution and Analysis: Imaging and scoring for 4 probe assay, 200 cells counted per sample	$7,775.00
DGH-004	in-Site Assay Execution and Analysis: Imaging and scoring for 3 probe assay, 200 cells counted per sample	$5,742.00
DGH-005	in-Site Assay Execution and Analysis: Imaging and scoring for 2 probe assay, 200 cells counted per sample	$4,642.00
DGH-006	in-Site Assay Execution and Analysis: Imaging and scoring for 1 probe assay, 200 cells counted per sample	$3,542.00
DGH-007	in-Site Custom Probe Production: Design and verification of custom probes	$5,500.00
DGH-008	in-Site Adherent/Suspension Metaphase Prep and Harvest	$1,000.00
DGH-009	in-Site T-Cell Culture Development: Thaw, recovery, and harvest optimization	$1,200.00
DGH-010	in-Site IPSC Cell Culture Development: Thaw, recovery, and harvest optimization	$1,300.00
DGH-011	in-Site Whole Blood Culture Development: Thaw, recovery, and harvest optimization	$900.00
DGH-012	in-Site T Cells Metaphase Prep and Harvest	$1,200.00
DGH-013	in-Site IPSC Metaphase Prep and Harvest	$1,300.00
DGH-014	in-Site Whole Blood Metaphase Prep and Harvest	$900.00
DGH-015	in-Site Assay Execution and Analysis: Imaging and scoring for 4 probe assay, 20 cells counted per sample	$1,166.25
DGH-016	in-Site Assay Execution and Analysis: Imaging and scoring for 4 probe assay, 50 cells counted per sample	$2,429.69
DGH-017	in-Site Assay Execution and Analysis: Imaging and scoring for 4 probe assay, 20 cells counted per sample with two quadrants per slide	$758.06
DGH-018	in-Site Assay Execution and Analysis: Imaging and scoring for 4 probe assay, 50 cells counted per sample with two quadrants per slide	$1,579.30
DGH-019	in-Site Assay Execution and Analysis: Imaging and scoring for 4 probe assay, 20 cells counted per sample with four quadrants per slide	$492.74
DGH-020	in-Site Assay Execution and Analysis: Imaging and scoring for 4 probe assay, 50 cells counted per sample with four quadrants per slide	$1,026.54

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dGH in-Site™

Examine Edits One Cell at a Time

1/4 -Example of in-Site and SCREEN combination assay in unedited human fibroblast control line cell. Bracketing in-Site probes on chromosome 22 (circled) to identify insert target location and dGH SCREEN paints on chromosomes 1, 2, and 3 (pink). No insert would be expected in this control line

3/4 - Example of the same in-Site assay in an edited iPSC line cell with no off target insertions. Target site bracketing probes in green and insert probe in yellow zoomed in and expanded on the right. Results from this sample indicated that 6% of cells had no off-target insertions.

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