Multiplex gene editing: engineer complex biology
Polygenic disease modeling
Recreate Complex Clinical Profiles
Most diseases are not driven by a single mutation. Simultaneously edit multiple genes to accurately model complex cancers, neurodegeneration, and metabolic disorders in vitro.
Pathway interrogation & drug resistance
Reveal True Phenotypes for Target ID
Single knockouts often show no effect due to cellular compensation. Knock out multiple paralogs or entire gene families at once to uncover true biological functions and validate novel drug targets.
Synthetic lethality and interactions
Discover Novel Therapeutic Vulnerabilities
Rapidly edit specific gene pairs to identify synthetic lethal interactions. Perfect for discovering new oncology targets and validating combination treatment strategies.
Precision delivery for complex multiplex edits
FluidFM delivers defined amounts of CRISPR reagents directly into individual cells, giving you greater control over editing conditions for complex multiplex projects.
By avoiding the harsh conditions of bulk transfection, it helps preserve cell health, improve editing specificity, and support more reliable downstream data.
Preserve cell integrity
Avoid the harsh conditions of bulk transfection and maintain the physiological and morphological state of your cells.
Improve editing specificity
Reduce over-dosing and off-target effects through controlled reagent concentrations.
Control reagent delivery
Deliver precise amounts of CRISPR RNPs and donor templates into each individual cell.
Protect data quality
Generate more reliable edited models by minimizing delivery-related stress and unwanted biological variability.
FROM DESIGN TO DELIVERY, WE HANDLE THE ENTIRE PROCESS
CellEDIT Workflow
Every CellEDIT project is designed, executed, and validated with maximum control and transparency. The strategy is designed by us to reduce your project time as much as possible while keeping cell integrity
Consultation
Project feasibility and CRISPR design strategy
Cell Pickup
Cell Packaging & Collection
CRISPR Design
Development of an editing strategy
CRISPR Injection
FluidFM delivery of CRIPR RNPs into single cells
Clonal Outgrowth
Single cell seeding and monoclonal outgrowth
Screening
Genotypic screening of targeted edits
Quality Control
Monoclonality, viability, sterility, and mycoplasma testing
Shipment
Cryopreserved clones and project report delivery
Included in every multiplex project
Every CellEDIT multiplex project includes the essential steps for design, editing, validation, and delivery of engineered cell lines.
| Deliverable | Included | On Demand |
| Editing strategy design from CellEDIT experts | ✓ | — |
| Clear project plan and regular updates | ✓ | — |
| Full gRNA design report before workflow begins | ✓ | — |
| Two edited clones & two Wild Type clones (1 cryovial per clone) | ✓ | — |
| Full final report: sequencing results, monoclonality proof, mycoplasma testing | ✓ | — |
| Full support with inbound and outbound cell shipment | ✓ | — |
| Additional clones | — | ✓ |
| Off-target analysis & report | — | ✓ |
Extended quality control (e.g. NGS, WGS, target copy number va) | — | ✓ |
Additional information for further analysis or publication | — | ✓ |
Multiplex gene editing FAQs
Find answers about multiplex editing, compatible cell lines, validation, and project requirements.
FluidFM technology uses hollow cantilever probes with nanoscale apertures to inject pre-assembled CRISPR ribonucleoprotein (RNP) complexes directly into the nucleus of individual cells.
Multiple guide RNAs targeting different genomic loci are co-delivered in a single injection step, enabling simultaneous editing of multiple targets without viral vectors or chemical transfection agents.
This has been demonstrated through successful triple knockout of FUT8, BAX, and DHFR in CHO cells in a single workflow.
Conventional transfection methods (lipofection, electroporation, viral transduction) deliver CRISPR components to the cytoplasm, where they must overcome nuclear import barriers — a bottleneck that reduces efficiency and makes equimolar multi-gRNA delivery unreliable.
Intranuclear injection via FluidFM bypasses the plasma membrane, cytoplasm, and nuclear envelope entirely, placing CRISPR components directly at the site of editing.
This eliminates cytoplasmic degradation losses, removes the need for viral vectors, and enables precise volume and concentration control across all target gRNAs simultaneously.
CellEDIT supports a broad panel of cell lines including CHO, U2OS, HeLa, MCF-7, C2C12, HEK293, MDA-MB-231, A549, and KPC cells. The service also accepts user-supplied cell lines, including hard-to-transfect primary cells and specialized research models that are difficult or impossible to engineer using conventional transfection approaches. No specialized transfection equipment is required from the customer.
The CellEDIT methodology for multiplex editing is validated in a peer-reviewed publication: Antony JS, Herranz AM, Gol TM, et al., "Accelerated generation of gene-engineered monoclonal CHO cell lines using FluidFM nanoinjection and CRISPR/Cas9," *Biotechnology Journal*, 2024.
The study demonstrates successful simultaneous triple knockout of FUT8, BAX, and DHFR in CHO-K1 cells using FluidFM intranuclear injection. Post-editing cell characteristics were not impaired, and the workflow was validated under serum-free media conditions compatible with biopharmaceutical manufacturing standards.
Multiplex editing accelerates pathway-level studies, synthetic lethality screens, epistasis analysis, and complex disease modeling requiring multiple mutations.
Biopharmaceutical applications benefit from simultaneous glycoengineering modifications (FUT8 knockout) and apoptosis resistance (BAX knockout) in production cell lines.
Drug discovery programs use multiplex editing for comprehensive target validation studies.
Monoclonality is guaranteed through FluidFM single-cell manipulation. Each edited cell line derives from one individually targeted cell that received the complete multiplex payload.
This eliminates the clonal heterogeneity inherent in bulk transfection methods where different cells in a population may receive different editing outcomes, requiring extensive screening.
Multiplex editing is available for CHO, U2OS, HeLa, MCF-7, C2C12, HEK293, MDA-MB-231, A549, and KPC cells. The service also accepts user-supplied cells. FluidFM intranuclear injection is particularly effective for hard-to-transfect cell lines where conventional multiplex approaches fail due to poor co-delivery efficiency.
Off-target effects are minimized through careful gRNA design with computational prediction for each target, vector-free RNP delivery that provides transient CRISPR activity, and direct nuclear injection that controls precise payload delivery.
The CellEDIT workflow includes comprehensive Sanger sequencing validation at all target loci to confirm intended edits and verify absence of off-target mutations.
Multiplex editing reduces timelines by months compared to sequential approaches. All target genes are modified in a single workflow, eliminating iterative cycles of editing, validation, and re-engineering.
This approach is essential for pathway-level studies where gene interactions must be assessed in the same genetic background. Guaranteed monoclonality from single-cell manipulation ensures genetic uniformity across all edits.
Yes, FluidFM intranuclear injection is particularly suitable for delicate and hard-to-transfect cell lines including primary cells and iPSCs. The gentle, vector-free delivery preserves viability where conventional electroporation or lipofection methods fail or cause excessive cytotoxicity.
FluidFM multiplex editing can target multiple genes simultaneously in a single injection. The CellEDIT service has validated triple knockout editing (FUT8, BAX, DHFR) in CHO cells.
The practical limit depends on gRNA design complexity and cell type, but the direct intranuclear delivery method supports co-injection of multiple RNP complexes without the efficiency losses seen in transfection.
Your Cells & Edits. Our Expertise.
Browse our custom cell lines: from hard-to-transfect to breast cancer cells, explore our CRISPR-edited cell lines through intra-nuclear delivery.
