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Custom Cell Lines Services 
CRISPR/Cas9 technology has been used as a genome editing tool for a variety of cell lines, including a variety of tumor cells in humans and mice. In addition, based on its easy operation and high accuracy, CRISPR/Cas9 technology have great potential for application in vitro disease models and disease treatment：
- CRISPR/Cas9-mediated fusion genes (chromosomal variants) for tumor researches.
- CRISPR/Cas9-mediated gene modification in primary T cells promoting the development of CAR-T immunotherapy.
- CRISPR/Cas9-mediated genetic modification in iPSCs providing disease-specific models for drug discovery.
CRISPR/Cas9 PlatformCB, the global leading biotechnology company focused on gene editing, provides a custom gene edited cell line service based on our self-successful CRISPR/Cas9 cell line generation platform. As one of the earliest CRISPR service providers, so far we have designed >1300 unique cell line models in >500 different mammalian cell lines. We can generate a CRISPR cell line model based on your specifications, which takes only 2-3 months.
Different CRISPR-mediated modifications we offered
| Frameshift mutation | Small nucleotide insertions or deletions (insert deletions) are introduced into the selected exons using a non-homologous end joining (NHEJ) repair pathway resulting in a frameshift of the ORF of the target gene, thereby altering the stop codon, which can lead to abnormally short or long changes in protein or amino acid sequence after translation, resulting in protein abnormalities. |
| Fragment excision | Using two gRNAs, large target regions of the gene of interest can be deleted, resulting in complete loss of function. Fragment excisions/deletions are useful for protein-coding and non-coding portions of the genome (e.g. promoter and enhancer regions, regulatory elements, etc.). |
| Insertion | Using a validated gRNA, Cas9 and donor DNA, short fragment DNA or transcription terminator signal is inserted into exons via homology directed repair (HDR) pathway. |
| Double gene knockout | Knockout of two or more genes is used to generate a multi-genetically modified cell line model. |
| Gene fusion | Generating cell line models with gene fusion (e.g. chromosomal translocation, deletion, inversion) are very clinically representative research models. |
The workflow of custom CRISPR/Cas9 cell lines generation
- Cell preparation.
Depending on the type of cell required, we prepare the cells two weeks in advance to maintain their high activity.
- Identification of target gene knockout genes.
Based on the nature of the gene, candidate knockout sites are selected and then identified.
- gRNA design and vector construction.
Three to five (3-5) sets of gRNA will be designed and construct the corresponding plasmids, while Cas9 is integrated.
- Transfection of cells and acquisition of monoclonal cells.
The constructed plasmid is transfected into a host cell to produce a cell bank. We will then sequence the cells in the cell bank to verify the knockout effect.
- Screen positive clones.
Transfected cells are seeded in 96-well plates for monoclonal screening using limiting dilution methods. Positive monoclonal cells will be amplified, sequenced, identified and stored. Finally, a cell line is successfully constructed.
- Stability test.
Mycoplasma detection and quantitative polymerase chain reaction and Western blotting.
- Cell line freezing and delivery.
Applications
- Physiologically relevant disease models that are difficult to mimic disease (e.g. amyotrophic lateral sclerosis, muscular dystrophy, Parkinson's disease, Alzheimer's disease).
- Differentiating mutations in different tissue lineages using isogenic cell line models.
- Suitable for targeted drug discovery, drug and toxicity screening.
- Molecular biomarkers for cancer prognosis research.
- Immunotherapy research.
Expected turnaround time
2~3 months
CRISPR/Cas9 PlatformCB is specializing in building efficient systems and procedures to meet the needs and timelines of clients working in the CRISPR/Cas9 gene editing. Our scientists have extensive experience with different cells, including easy-to-transfer cell lines and difficult-to-transfect cell lines. Tell us your project needs, we will offer you a one-stop-shop CRISPR/Cas9 gene editing service from strategy design to final stable cell line. We guarantee our clients the most reliable and efficient research services to best match your research goals, and faster turnaround time, lower price. If you have any questions, don’t hesitate to contact us.