Immortalized Cells: Enhance Your Research with EditCo's Reliable CRISPR-edited Cell Lines for High-Quality Reproducible Results

Access CRISPR Cell Lines Without Sacrificing Time

EditCo’s Knock-in Immortalized Cells are your quickest path to integrating engineered cells into your research. Whether modeling disease with SNVs or tagging proteins for in-depth studies, DIY CRISPR projects often come with unforeseen challenges and delays, typically requiring several experimental repeats and adding months to your timeline. EditCo’s advanced automated transfection optimization and top-tier reagents maximize editing efficiency, guaranteeing a successful CRISPR edit.

Unlock your research potential with EditCo’s Knock-in Immortalized Cells and accelerate your path to discovery.

Successful Knock-ins in Any Cell Line

EditCo’s CRISPR Knockout Cell Clones eliminate the time and hassle of doing CRISPR editing and creating clonal cell lines so you can focus on your research.

Figure 1. Successful knock-in editing, even in hard-to-edit cell lines, enabled by our automated transfection optimization protocol. gRNAs and ssODNs were designed to induce a 33nt knock-in and then transfected into the 9 different immortalized cell lines indicated. Post recovery, knock-in editing efficiencies were measured in the mixed population of edited and unedited cells using EditCo’s ICE Knock-in analysis tool. The cells are then either used directly for assays or isolated into clonal populations.

Figure 2. High knock-in efficiencies across numerous loci. sgRNA and ssODNs were designed to induce a 9-nt knock-in at numerous different loci, then transfected into multiple different cell types. Post cell recovery, knock-in editing efficiencies were assessed using EditCo’s ICE Knock-in analysis tool. The values displayed above represent the percentage of each total population of cells that have the desired knock-in edit present.

Generate Assay Results with Speed and Confidence

Express Cell Pools are highly knockout-edited pools of cells consisting of a heterogeneous population of edited and unedited cells for your gene of interest in human and mouse immortalized cell lines, generated in 2 weeks or sooner.

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 Express Cell Pools are the solution for obtaining quick and reliable functional data while reducing the risk for false negatives in your critical assays.

Express Cell Pools workflow includes the use of our unique multi-guide design, transfection, genotyping, and sequencing analysis. Choose from either EditCo supplied cell lines banked from our high-quality cell supplier ATCC, or onboard your cells.

Alternatively, if you would like to use your sgRNA sequence, Express Custom Cell Pools are also available. These allow user-defined sgRNA sequences to specifically target the transcript or genomic region of interest. Please contact us for further information.

Available ATCC Cell Lines for Express Knockout Cell Pools

If the cell line you are interested in is not available in above table, learn about onboarding your own below. 

EditCo's Express Cell Pool Service for Your Unique Cell Lines

Whether you're in need of human tumor cell lines from esteemed sources like the National Cancer Institute, your trusted workhorse cell line, or a previously edited line, EditCo is dedicated to simplifying your research journey. Bid farewell to the complexities of cell optimization and gene editing–simply onboard your cells, and let us take care of the cell editing with our state-of-the-art Express Cell platform.

Our User-Supplied Express Cell Knockout Pools involve a straightforward two-step process:

1. Cell Banking: Your supplied cell line undergoes EditCo's proprietary banking process, ensuring high quality through quality control (QC) measures. This includes a strict QC to ensure we can deliver high-quality edited cells in 2 weeks or less after the cell banking is complete. A detailed QC report is provided with all the parameters evaluated.
   
   
2. Cell Editing: Once successfully banked, up to 20 edits can be generated per banking cycle. You have the flexibility to choose when edits are performed, with no minimum requirement. Our edited cell knockout pools guarantee at least 80 percent editing efficiency. Edited cells undergo the same rigorous QC process as our standard Express Cell platform. Each edit includes 2 vials of edited cell pools and 2 vials of wild-type cells, each containing over 300,000 cells per vial.
   
   

Key Benefits:

• Flexibility: Perform up to 20 edits from a single banking cycle.
• Fast turnaround: Receive edited cell pools (>300,000 cells/vial) in 2 weeks or less.
• High efficiency: We guarantee 80 percent editing efficiency for all knockout pools.
• Quality assurance: We perform stringent QC processes for banking and editing.
  
  

*Cell lines must meet the Express Platform growth conditions: < 72 hrs doubling time, no coating or shaking required, no complex media requirements, ability to grow at 37 C, 5% CO2, and BSL-2 safety only.

Figure 1. Express Cell Pools generated across 355 genes and 36 different cell lines in just 12 days. Using EditCo’s multi-guide designs, up to 3 modified gRNAs per target were conjugated with SpCas9, and RNPs were transfected into cells. On average, Express Knockout Cell Pools were generated in 12 days and achieved >80 percent Gene Knockout (Knockout Score percentage).

Sustained Gene Knockout Across Cell Passages

Figure 2. High editing efficiency is sustained through multiple cell passages. Using EditCo’s multi-guide designs, up to 3 modified gRNA per target (across 6 genes) were conjugated with SpCas9, and RNPs were transfected into U2OS cells. Editing efficiencies were measured at different time points post-transfection and up to 4 passages. The median editing efficiency is >85 percent for each of the targets. 

Cell Viability Is Maintained Across Passages

Figure 3. Cell viability is maintained through multiple cell passages in U2OS cells. Viability was taken using Celigo. Please note that CDK9 is a common essential gene, hence the lower viability and cell number observed along the passages.

Our Unique Guide Design Powers Functional Knockouts and Sustained Protein Depletion

Figure 4. The multi-guide approach consists of 3 gRNAs spatially coordinated to create a fragement deletion in a single exon. Targeting a single early exon of a gene induces multiple concurrent double-strand breaks in the target sequence of a gene. This results in disruptive one or more >21 bp fragment deletions. This approach is associated with higher frequencies of knockout alleles in edited pools but reduced occurrence of off-target edits as compared to single-guide approaches.

Figure 5. Edited cell pools utilizing the multi-guide technology shows persistent protein depletion. This enables direct and reliable use of these cell pools for functional assays. Western blot analyses for all 5 knockout target genes indicate a complete depletion of proteins across the 3 specified time points (days 7, 14, and 21) relative to the negative controls. Knockout cell pools for the target genes were generated by nucleofecting U2OS cells with multi-guide sgRNA and Cas9 (as RNPs). A negative control pool was also transfected for each target using non-targeting sgRNA.

Validate Drug Targets with Speed and Confidence

The high-throughput arrayed CRISPR screening format has emerged as a popular approach for target discovery because the results are amenable to straightforward interpretation without requiring labor-intensive data deconvolution steps and compatibility with a wide assortment of downstream assays. However, this approach has historically remained underutilized due to lack of infrastructure, challenges associated with transfection optimization, and/or limited bandwidth.

EditCo’s Engineered Cell Libraries lower these barriers to broaden the accessibility of CRISPR as a drug target discovery tool.

Engineered Cell Libraries are custom panels of edited immortalized or iPS cell pools in an arrayed format suitable for direct application in functional assays. EditCo's smart multi-guide design strategy enables editing efficiencies typically higher than 90 percent. Combined with the power and cell engineering capabilities high-throughput editing platform, you can generate libraries at any experimental scale, start your screen quickly, and confidently select the best hits.

Streamlined Intake and Optimized Process Yield Predictable Knockouts

The Engineered Cell Libraries workflow includes multi-guide design, high-throughput transfection, knockout pool generation, genotyping, and sequencing analysis.

Knockouts in Human Immortalized Cell Line

Figure 1. A library of knockout cell pools corresponding to 42 gene targets was generated via high-throughput editing in human cells. Data from the experiment are shown above. An Engineered Cell Library with 42 gene targets was generated via high-throughput editing of an immortalized-adherent human cell line, A375, using multi-guide technology. The data demonstrate both indel frequencies and Knockout Scores. Of the 84 percent of pools successfully sequenced and analyzed, an average indel frequency of 99 percent and an average Knockout Score of 98 percent were observed.

Knockouts in Mouse Immortalized Cell Line

Figure 2. A library of knockout cell pools corresponding to 192 gene targets was generated via high-throughput editing in mouse cells. The data demonstrate both indel frequencies and Knockout Scores. Among the 80 percent of pools successfully analyzed, the average indel frequency was 89 percent, while the average Knockout Score was 84 percent.

Our Unique Guide Design Powers Functional Knockouts and Sustained Protein Depletion

The multi-guide approach consists of up to 3 gRNAs (green bars) spatially coordinated that target a single early exon of a gene and induce multiple concurrent double-strand breaks in the target sequence. This results in disruptive one or more >21 bp fragment deletions. This approach is associated with higher frequencies of knockout alleles in edited pools but reduced occurrence of off-target edits as compared to single-guide approaches.

Figure 3. The multi-guide approach consists of 3 gRNAs spatially coordinated to create a fragement deletion in a single exon. Targeting a single early exon of a gene induces multiple concurrent double-strand breaks in the target sequence of a gene. This results in disruptive one or more >21 bp fragment deletions. This approach is associated with higher frequencies of knockout alleles in edited pools but reduced occurrence of off-target edits as compared to single-guide approaches.

Figure 4. Edited cell pools utilizing the multi-guide technology shows persistent protein depletion. This enables direct and reliable use of these cell pools for functional assays. Western blot analyses for all 5 knockout target genes indicate a complete depletion of proteins across the 3 specified time points (days 7, 14, and 21) relative to the negative controls. Knockout cell pools for the target genes were generated by nucleofecting U2OS cells with multi-guide gRNA and Cas9 (as RNPs). A negative control pool was also transfected for each target using non-targeting gRNA.