CRISPR Stable Knockout Cell Line Service
Cat. No. C208
When precision matters, trust abm, a global leader in custom cell line engineering. With extensive CRISPR expertise and hundreds of successful projects delivered worldwide, we provide seamless, reliable generation of stable knockout cell lines tailored to your research goals.
Using a robust CRISPR/Cas9 lentiviral system, we generate stable knockout cell lines in human, mouse, rat and canine models, optimized for your specific cell type and experimental requirements. Our platform has been successfully applied across a wide range of cell types, including fibroblasts, blood-derived cells, ovarian cells, hepatocytes, stem cells, and more.
Simply provide your gene of interest and cell culture conditions — we handle the entire workflow.
Comprehensive End-to-End Workflow
- sgRNA design and optimization
- Molecular cloning and construct validation
- Lentiviral packaging
- Transduction or transfection and antibiotic selection
- Isolation and expansion of stable knockout clones
- Genotype validation
Why Choose abm?
- Proven CRISPR platform with versatile delivery options (plasmid or viral)
- High editing efficiency and reproducibility
- Experience across diverse cell types
- Dedicated technical support from project initiation to delivery
Choose Your CRISPR KO Service Level
| Base | Standard ★ | Premium | |
|---|---|---|---|
| Validated KO Clones | 1 Clone | 2 Independent Clones | 3 Independent Clones |
| Starting Price | $1,690 | $2,590 | $3,490 |
| sgRNA Design | ✓ | ✓ | ✓ |
| Vector/Virus Construction | ✓ | ✓ | ✓ |
| Stable Cell Line Generation | ✓ | ✓ | ✓ |
| Sequencing + ICE Analysis | ✓ | ✓ | ✓ |
| Sterility / Mycoplasma Test | ✓ | ✓ | ✓ |
| Experimental Confidence | Basic | High | Maximum |
| Recommended For | Proof-of-concept | Publication-ready studies | High-impace/reproducibility-critical studies |
Deliverables
- 2 Frozen Vials per Clone
- Service Report
Customer Requirements
The customer must provide:
- A minimum of 2 million viable cells unless customer opts to select from abm’s cell service library
- 1 L of propagation media*
*If the cell line uses standard DMEM or RPMI, abm will supply the media.
- Any required specialized coated flasks, if applicable
The submitted cell line must:
- Tolerate single-cell cloning
- Demonstrate adequate transfection and/or transduction efficiency
Have questions? Connect with our team at inquiries@abmgood.com.
Add-On Services
For proper experimental controls, be sure to request a Wild Type Control Cell Line expressing Cas9—ensuring reliable comparison within the same cellular background.
Enhance your CRISPR knockout project with optional validation and customization services designed to strengthen your experimental outcomes.
| Add-On Services | Unit | Cat. No. | Price |
|---|---|---|---|
| WT Control Cell Line Expressing Cas9 for Comparison | 1 Cell Line | C142 | $1590.00 |
| 50% discount for academic customers if ordered with service | |||
| Additional Clones^^^ | 1 Clone | C143 | $900.00 |
| Additional Vials of Delivered Cells^^^ | 1 Vial | C144 | $300.00 |
| Western Blot Validation Service ## | 1 Clone | C150 | $2000.00 |
| Cell Line Authentication: STR Profiling for Human samples | 1 Sample | C287 | $195.00 |
| For other species, please inquire | |||
Notes:
Please note that if the gene to be knocked-out may be essential to cell survival, it is up to the end-user to proceed with the services. abm is unable to guarantee cell survival in these cases and will only attempt to rescue the clones under these conditions. abm is not accountable for cell survival if rescuing the clones (instructions to be provided customers) is unsuccessful.
## For Western blot validation services, customer must provide pre-validated antibody, along with an appropriate positive control (preferably with supporting data).
^^^ Additional media, supplements, or optimization steps may be required depending on cell type.
CRISPR Stable KO Service Case Studies
Customer Review:
"We are quite happy with the knockout cells that you made. This was a tricky knockout and hemizygous removal was much more prevalent than the double knockout, but you did it. In addition to your excellent genomic sequence characterization of both mutant LIF alleles, I have confirmed in our lab that the tumor cells have lost all mouse LIF production."
Dr. Robert Jackman, Boston University, Mouse LIF CRISPR Knock Out C26 Tumor Cell Line Generation and Screening Service, View case study here.Additional Resources:
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FAQs
| What should customers provide for this service? |
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Please supply the cells for this service. For frozen cells, please ship at least 2 vials (at least 106 each) on dry ice. An alternative method is to send us two T25 flask of live cells per sample, at 90% confluency. The flasks should be filled with complete medium without any air bubbles and at room temperature. Please ensure that the sterilization procedure is strictly observed. To avoid delays over the weekend, we recommend shipping the cells on a Monday or a Tuesday. Frozen cells are preferred over live cells. If your cell line already contains an antibiotic resistance marker, please specify. Our vectors have puromycin resistance by default. If your cells require medium other than DMEM and RPMI, we will also require 1L of the specified medium and any applicable growth factors or supplements to be provided by you for the project. Please submit all components of the complete media (e.g. if growth factors, cytokines etc. are applicable) individually to eliminate potential degradation of components in the media during transit. Kindly include instructions for making the complete media in your shipment. Additionally, please provide at least 5 x coated 6-well plates and 10 x T25 flasks if the cells require specially treated culture vessels. Once you have shipped your cells, please forward us the tracking number for custom clearance. Information on how to ship cell samples to abm can be found on our support page: /Technical-Support.html Please place an order first prior to submitting your samples. All samples received must have the order confirmation number indicated. Any samples received without this piece of information will be disposed off immediately upon receipt to ensure that all customer information is held in strict confidence.
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| Can this service be performed for canine derived cells? |
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Yes, we can use sgRNA targeting canine genome to make stable KO cell lines. Please provide the specific cell medium for canine cell culture.
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| Will CRISPR keep cutting the chromosome after the gene is edited? |
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CRISPR is sensitive to mismatches, so it is unlikely the CRISPR will keep cutting the chromosome after the gene is edited.
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| Is Nuclease or Nickase used? Can you use the double Nickase approach? |
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The approach used for the above service is sgRNA with Cas9 Nuclease, relying on NHEJ repair without a repair template. The Nickase approach is available but this will be at an additional charge and will be subject to a custom quotation as a different strategy will need to be devised.
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| How can you detect a mutation if there is no selection (by PCR using primers flanking the modified site or the T7 endonuclease I assay)? |
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Yes, that is one method used. Other methods include Sanger sequencing, and Western Blot if the antibody is provided by the customer.
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| How many sgRNAs are designed and synthesized for this service? Are pooled sgRNAs used? |
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We design 3 sgRNAs at the time of order placement, however only 1 sgRNA is used to achieve knockout. In the case that the first sgRNA does not allow for success, we would try another from the set of 3 synthesized sgRNAs.
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| Can you offer off-target site prediction service and off-target site analysis service (Sanger sequence)? |
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When we design the sgRNAs, the design software will predict the number of off-targets, which is usually 0 for the sgRNAs that we select. Off-target analysis: This is typically done by NGS, whole genome sequencing or exome sequencing, which would require comparing the control (before) sample to the genome edited sample (after).
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| How is KO confirmed? |
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We perform Sanger sequencing of the locus of interest both to determine the nature of the INDEL in the edited genes to guarantee frameshift silencing, and to confirm that the genome is edited.
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Citations
| 01 |
Kandarian, Susan C et al. “Tumour-derived leukaemia inhibitory factor is a major driver of cancer cachexia and morbidity in C26 tumour-bearing mice.” Journal of cachexia, sarcopenia and muscle vol. 9,6 (2018): 1109-1120. doi:10.1002/jcsm.12346 |
| 02 |
Kang, Y. J. et al. "Regulation of NKT cell-mediated immune responses to tumours and liver inflammation by mitochondrial PGAM5-Drp1 signalling." Nat. Commun. (2015) 6:8371 doi: 10.1038/ncomms9371 |
| 03 | Jiang, G. et al. "Isorhapontigenin (ISO) inhibits invasive bladder cancer (BC) formation in vivo and human BC invasion in vitro by targeting STAT1/FOXO1 Axis." Cancer Prev Res. Published Online First April 14, 2016.doi: 10.1158/1940-6207.CAPR-15-0338 |
| 04 | Okugawa, Y. et al. "Clinical significance of SNORA42 as an oncogene and a prognostic biomarker in colorectal cancer." Gut (2015)doi:10.1136/gutjnl-2015-309359 |