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Primary Cell KO Pools: Advance your immunology research with edited-to-order primary cells

High editing efficiency with reliable post-editing functionality. Order Primary Cells supplied by EditCo or work with our team to edit your primary cell type of interest.

Using other primary cell lines? Reach out to us about custom projects!
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Overview

Efficient edits, functional T-cells, better results

  • Confident Knockouts: 80% editing efficiency guaranteed with our 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 CD8+ T-cells.
  • Functionality: High cell viability and editing levels without affecting cytotoxicity.
T-cell image

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.

EC Pools overview-1M cells

 

Want to learn more? Click here to schedule a free consultation with a CRISPR expert today!

Contact Us

Deliverables

Focus on your experiment and leave CRISPR to us.

Our KO Primary T-cells are guaranteed to have at least 80% editing efficiency. Now available using EditCo supplied cell lines or onboard cells from your experiments or clinical cohorts.

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.

Contact Us

Data

Stability and Functionality Post-Editing

High and Stable Editing Efficiency of CD8+ T-cell pools. 

EC_KOPool-Stability

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.

CD8_T-cell_viability-1

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

CD8 MFI grouped final

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

CD8 activation final

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

Flyer
Knockout CD8+ T-cell Pools
Edited-to-order Knockout CD8+ T-cells guarantee high knockout efficiency while retaining high cell viability and post-editing functionality.
Learn More
Poster
Rapid and Efficient Genome Editing for Human Primary T-Cells
EditCo can quickly provide functional, stable, made-to-order edited T-cells from a variety of prevalidated donors using our genome engineering platform. Presented at Immuno-Oncology 2024.
Learn More
Flyer
Industrialized CRISPR Cells: CRISPR Done for You
EditCo's next-generation engineered cells provide researchers with access to the benefits of CRISPR. You no longer need to invest significant time and money to learn and optimize methods.
Download Now
Peer-reviewed Paper
Non-viral expression of chimeric antigen receptors with multiplex gene editing in primary T-cells
Learn how we're carving a path for potential "off-the-shelf" CAR-T therapies in collaboration with the Saha lab at the University of Wisconsin - Madison.
Learn More
Overview

Skip to the main experiments with edited CD4+ T-cells

  • 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.
Untitled-1

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.

EC Pools overview-1M cells

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!

Contact Us

Deliverables

Focus on your experiment and leave CRISPR to us.

Our KO Primary T-cells are guaranteed to have at least 80% editing efficiency. Now available using EditCo supplied cell lines or onboard cells from your experiments or clinical cohorts.

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.

Contact Us

Data

Stability and Functionality Post-Editing

Custom-edited CD4+ T-cells

EC_edit-efficiency

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).

CD4KO flow panel

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).

CD4_data_web

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.

CD4_secretion_web

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

Poster
Rapid and Efficient Genome Editing for Human Primary T-Cells
EditCo can quickly provide functional, stable, made-to-order edited T-cells from a variety of prevalidated donors using our genome engineering platform. Presented at Immuno-Oncology 2024.
Learn More
flyer
Knockout Primary Immune Cell Pools
Edited-to-order Knockout CD4+ T-cells guarantee high knockout efficiency while retaining high cell viability and post-editing functionality.
Learn More
Peer-reviewed Paper
Non-viral expression of chimeric antigen receptors with multiplex gene editing in primary T cells
Learn how we're carving a path for potential "off-the-shelf" CAR-T therapies in collaboration with the Saha lab at the University of Wisconsin - Madison.
Learn More
Flyer
Industrialized CRISPR Cells: CRISPR Done for You
EditCo's next-generation engineered cells provide researchers with access to the benefits of CRISPR. You no longer need to invest significant time and money to learn and optimize methods.
Learn More
EditCo's Automated CRISPR Platform

Integrating our core CRISPR expertise, high-quality reagents, and automated processes, we deliver the best edited cell-based models at any scale.

CRISPR Engineered Cells Enable Variant Disease Modeling
Icon_Pools_Trans
Leveraging a sophisticated infrastructure with integration between bioinformatics, software, and automated platforms
Learn More
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Automating biology and synergistic disciplines for streamlined cell editing and cell culture workflows
Learn More
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Delivering experiment-ready edited cells for your next discoveries through a robust and cohesive ecosystem
Learn More
Testimonials 1

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Principal Investigator, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School
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Principal Investigator, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School
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Albert Rose, Ph. D.
Principal Investigator, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School
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Albert Rose, Ph. D.
Principal Investigator, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School
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