Efficient edits, functional T-cells, better results
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Confident Knockouts: 80% editing efficiency guaranteed with our smart multi-guide design.
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Faster Results: A 7-day editing protocol delivers results in 2 weeks or faster
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Flexibility: Choose from EditCo-supplied cells or onboard your CD8+ T-cells.
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Functionality: High cell viability and editing levels without affecting cytotoxicity.
Get Guaranteed Editing Results in Primary Cells
Primary Cell KO Pools are made-to-order knockouts with a guaranteed editing efficiency of >80% in human primary cells generated in 2 weeks or faster. EditCo consistently achieves >90% editing efficiency, creating cell pools that are functionally similar to a clone. As primary cell clones are biologically difficult to produce, EditCo’s high editing efficiency provides a unique approach to generating cell populations of interest.
Whether you are leveraging CRISPR for validating targets for drug discovery or modeling a biological process for the investigation of a novel gene of interest, failed experiments lead to a loss of time and resources. EditCo’s Primary Cell KO Pools are the solution for obtaining quick and reliable functional data while reducing the risk for false negatives in your critical assays.
Want to learn more? Click here to schedule a free consultation with a CRISPR expert today!
Focus on your experiment and leave CRISPR to us.
Features
Cell Source
- EditCo supplied
- Customer supplied
Available Edits
- Single knockout
CRISPR Design
- Multi-guide synthetic modified sgRNA (standard)
Deliverables
- Regular updates on your order's progress
- 2 vials of edited cell pools with >1,000,000 cells/vial (pools consist of a heterogeneous population of edited and unedited cells)
- Control-transfected cell pools (2 vials)
- Sequence of synthetic gRNA used
- Primer sequences used for PCR and Sanger Sequencing
- ICE Sanger sequencing analysis reports for each edited clone after expansion
- Comprehensive QC report that includes the following information: mycoplasma test (positive/negative) and passage number
Other Primary Cells
EditCo is expanding its Primary Cell portfolio. Please reach out to inquire about additional cell types, including primary fibroblasts, monocytes/macrophages, NK cells, B cells and primary mouse cell types.
Stability and Functionality Post-Editing
High and Stable Editing Efficiency of CD8+ T-cell pools.
Figure 1. High editing efficiency across multiple donors. Three different donors (A, B and C) show high editing efficiency (>90%), as determined by ICE analysis, across various targeted loci with little change between deliverable (before cryopreservation, left graph) and extended culture (28 days post-thaw, right graph). Cell stimulation after thawing did not affect KO levels.
High viability and expansion of edited pools
EditCo KO CD8+ T-cell pools can be thawed and expanded for weeks, with or without TCR stimulation.
Figure 2. CD8+ viability and expansion after thaw. Edited CD8 T-cell pools were thawed and plated in G-RexTM-24 consumables at 1E6 cells per well in ImmunocultTM T-cell expansion media supplemented with 100ng/ml recombinant human Interleukin-2. Stimulated samples were treated with ImmunocultTM CD2-CD3-CD28 T-cell activator for 3 days. Cultures were fed every 3 days by replacing 90% volume with fresh media. On Day 14* cell number was returned to 1E6 cells per well to prevent overgrowth.
Biologically Relevant
Figure 3. Preserving cell function. EditCo T-cell pools undergo a single round of activation to preserve normal cell function and prevent the cells from acquiring an overstimualted or exhausted phenotype. This is readily seen by the transient expression of activation markers (CD25, CD69) and exhaustion markers (PD1).
Antigen-specific T-cell Activation
Figure 4. T-cell activation measured by CD107a. Edited CD8+ pools were stimulated with mixed viral peptides (CEF, REF 3) presented on HLA-1 matched B-cells. After 2 rounds of stimulation T-cells were incubated with CEF peptides for 4 hours, and cytotoxic activation was measured by CD107a surface localization. WT, CD2, ICAM-2 edited pools showed robust CD107a expression compared to DMSO controls. As expected, B2M and TRAC knockout pools showed no induction above baseline, given their role in antigen presentation and recognition.
Resources
Skip to the main experiments with edited CD4+ T-cells
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Confident Results: 80% editing efficiency guaranteed smart multi-guide design.
-
Faster Results: A 7-day editing protocol delivers results in 2 weeks or faster
-
Flexibility: Choose from EditCo-supplied cells or onboard your CD4+ T-cells.
-
Functionality: High cell viability and editing levels without affecting functionality.
Get Guaranteed Editing Results in Primary Cells
Primary Cell KO Pools are made-to-order knockouts with a guaranteed editing efficiency of >80% in human primary cells generated in 2 weeks or faster. EditCo consistently achieves >90% editing efficiency, creating cell pools that are functionally similar to a clone. As primary cell clones are biologically difficult to produce, EditCo’s high editing efficiency provides a unique approach to generating cell populations of interest.
Whether you are leveraging CRISPR for validating targets for drug discovery or modeling a biological process for the investigation of a novel gene of interest, failed experiments lead to a loss of time and resources. EditCo’s Primary Cell KO Pools are the solution for obtaining quick and reliable functional data while reducing the risk for false negatives in your critical assays.
The Primary Cell KO Pools workflow includes the use of our unique guide design, transfection, genotyping and sequencing analysis. Choose from either EditCo supplied cell lines banked from our high quality third party suppliers or onboard your cells.
Want to learn more? Click here to schedule a free consultation with a CRISPR expert today!
Focus on your experiment and leave CRISPR to us.
Features
Cell Source
- EditCo supplied
- Customer supplied
Available Edits
- Single knockout
CRISPR Design
- Multi-guide synthetic modified sgRNA (standard)
Deliverables
- Regular updates on your order's progress
- 2 vials of edited cell pools with >1,000,000 cells/vial (pools consist of a heterogeneous population of edited and unedited cells)
- Control-transfected cell pools (2 vials)
- Sequence of synthetic gRNA used
- Primer sequences used for PCR and Sanger Sequencing
- ICE Sanger sequencing analysis reports for each edited clone after expansion
- Comprehensive QC report that includes the following information: mycoplasma test (positive/negative) and passage number
Other Primary Cells
EditCo is expanding its Primary Cell portfolio. Please reach out to inquire about additional cell types, including primary fibroblasts, monocytes/macrophages, NK cells, B cells and primary mouse cell types.
Stability and Functionality Post-Editing
Custom-edited CD4+ T-cells
Figure 1. Editing Efficiency and Viability for CD4+ T cell pools. Values represent the average of three different donors showing high editing efficiency, as determined by ICE analysis, across a variety of loci (blue bars, mean KO scores +- SEM) without affecting viability at the time of freezing (cyan dots).
Figure 2. Editing Efficiency leads to protein depletion as shown by flow cytometry (BD Symphony A1).
High viability and expansion of edited pools
The editing process and further expansion before freezing do not affect cell fitness. Edited cells can be thawed and expanded for any desired downstream assay, maintaining high cell viability and editing levels (Figure 3).
Figure 3. CD4+ pool stability after thaw. Panel A. PD1 expression in TCR activator-treated cultures peaked on day 3 in all tested donors (edited and mock samples). Isotype control staining over the same period is shown for reference. Panel B. Relative fold expansion for edited pools cultured with TCR activator or IL2 alone. Panel C. Pools of 4 different edits from 3 separate donors showed >90% viability and no decrease in editing efficiency after 14 days in culture.
High functionality
Edited CD4+ T-cells produced levels of IFNg, IL2 and TNFa after mitogenic stimulation that correlated with known target gene function.
Figure 4. Cytokine secretion in edited cell pools following cell expansion. Edited cells from 3 donors were thawed and then stimulated with PMA/ionomycin for 6 hours. Supernatants were harvested and measured for concentrations of IFNG (left panel), IL2 (center panel) and TNFA (right panel) as well as IL10 and IL5 (not shown) by FACS-ELISA (Miltenyi). Values were quantile normalized to reduce batch effects. Following ANOVA, Tukey’s multiple comparison test was performed between Mock and each gene target to determine statistical significance. (*) p<0.05; (**) p<0.01; (***) p<0.005; (****) p< 0.0001.
Figure 5. Intracellular Cytokine measurement in edited cell pools. Knockout CD4+ T-cell pools of IFNg, IL2 or TNFa were thawed and stimulated with PMA/ionomycin for 6 hours. Cells were stained for CD3, CD4 and viability, permeabilized and then stained for presence of IFNG, IL2 or TNFA. Production of either cytokine in CD3/CD4/Live gated events for each Gene Target knockout culture (Bottom histograms) was compared to unedited control (Mock, Middle histograms) as well as TRAC knockout (Top histograms).
Resources
Integrating our core CRISPR expertise, high-quality reagents, and automated processes, we deliver the best edited cell-based models at any scale.
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