CRISPR-GENERATED CELL-BASED MODELS
Enhance your insight into human disease with our CRISPR modeling service
Overview
When you can't find the right iPSC line or iPSC donor material to create a cell-based model for a specific disease, DefiniGEN can help. With our years of experience generating mature, primary-like cells from iPSCs and our proficiency at using CRISPR for targeted genome editing, we can build the cells that best reflect the biology you wish to model.
We can use CRISPR gene editing to generate cell models that:
- Repair causative mutations in a patient-derived line
- Introduce causative alleles into a healthy parental line
- Are isogenic with existing healthy or disease-state cells
- And more
Contact us to learn more >
Want to do the differentiation yourself? We can perform the CRISPR genome editing and send you the bespoke iPSCs
Learn more about our gene editing service >
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Rapid turnaround timeOur optimized protocols and standardized workflows ensure fast, efficient project completion. -
High success ratesOur deep expertise handling challenging iPSCs easily translates into successful CRISPR editing of iPSCs. -
Collaborative project managementWe conduct every project with complete transparency and in close collaboration with client teams to ensure we deliver what you need.
What we offer
Our custom cell model service is an integrated, end-to-end package that includes iPSC line or donor selection, the complete CRISPR editing workflow, and differentiation into the target cell type. From basic frameshift knock-out mutations to complex knock-in genetic changes, our dedicated scientific team will work collaboratively with you to design and build the most relevant cell-based model for your project.
Contact us to learn more about working with DefiniGEN - we respond within 1 business day to every inquiry.
We are an approved supplier on scientist.com
What you can expect
Our workflow consists of footprint-free CRISPR/Cas9-mediated editing, single-cell cloning of the edited population, and characterization of expanded clonal cell lines to identify positive clones. The edited iPSC cells maintain pluripotency and have normal, stable karyotypes, which are important for testing downstream applications such as directed differentiation.
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iPSC Reprogramming or SourcingWe can start your service with any of our comprehensively characterized in-house iPSC lines*, you can supply your own iPSC lines, or you can provide material derived from patient donors. We can even help you find the right donor material from our extensive clinical network. -
Gene editingWe work in collaboration with you to design sgRNAs and use a footprint-free CRISPR/Cas9 system to edit the iPSCs. We optimize transfection conditions before full-scale editing, and then screen the edited populations, seed single cells, and sanger sequence the clones. You select which clones to move forward with. -
Differentiation and bankingWe use our world-leading OptiDIFF platform to differentiate your edited iPSCs. Once the differentiation process is complete, we cryopreserve and bank the differentiated cells.OptiDIFF platform -
Product validationWe use a variety of analytical tools, including genotyping, phenotyping, and cell-based assays, to ensure that the target cells possess the expected characteristics of the mature cell. -
DeliveryUpon successful validation, we ship your cryopreserved differentiated cells to you, and you can expect high viability post-thaw.
*Using our in-house iPSCs can increase your chances of success as we already have a deep understanding of how these iPSCs will respond to CRISPR editing and during differentiation.
Disease Modelling
We can give your project a big head start with our well-characterized in-house iPSCs that model a number of monogenic diseases, many with multiple different models (Table 1 below).
When modelling any pathology using an iPSC differentiation platform, it is vital to confirm that the metabolic pathways implicated are expressed and active in the wild-type cells. DefiniGEN's hepatocytes are mature, predictive, and display all relevant biological pathways pertinent for our disease models, as these data show below.
Monogenic diseases we can model
| Monogenic Diseases | |
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Gaucher disease |
Acute intermittent porphyria |
| Genetic cholestasis (PFIC, TGP2, and Alagille syndrome) | Familial hypercholesterolemia |
| Wilson’s disease | Organic acidurias |
| Hereditary hemochromatosis | Cystic fibrosis |
| Tyrosinemia type 1 | Erythropoietic protoporphyria |
| Alpha-1 antitrypsin deficiency | MCAD deficiency |
| Arginosuccinic aciduria (ASL) | D-bifunctional protein deficiency |
| Glycogen storage disease (GSD) type 1 | Galactosemia Type 1 |
| Urea cycle disorders | Citrullinemia |
| Crigler-Najjar syndrome | Familial amyloid polyneuropathy |
| Primary hyperoxaluria type 1 | Atypical haemolytic uremic syndrome 1 |
| Maple syrup urine disease (MSUD) |
Wilson's Disease
The ATP7B gene is fundamental in hepatocytes for enabling the cells to effectively metabolise copper, and mutations in this gene can lead to Wilson’s disease. The ATP7B gene is shown here by qPCR to be expressed at comparable levels to human primary cells and the pathway is active in DefiniGEN’s differentiated hepatocytes, which make a good isogenic control for our disease model cells.
Gaucher's Disease
Gaucher’s disease is caused by the defective activity of the lysosomal hydrolase glucocerebrosidase, which is encoded by the GBA gene. The GBA pathway is present in DefiniGEN’s differentiated hepatocytes, shown here by qPCR, which provide a good isogenic control to our disease model cells.
Crigler-Najjar Syndrome
Crigler-Najjar syndrome is a rare monogenic disorder which is caused by a mutation in the UGT1A1 gene. The UGT1A1 enzyme is required for the conjugation and excretion of bilirubin from the body. The UGT1A1 pathway is shown here by qPCR to be expressed sufficiently in DefiniGEN’s wild type differentiated hepatocytes for them to be used as isogenic controls to our disease model cells.
Familial Transthyretin Amyloidosis
Familial Transthyretin Amyloidosis is a rare inherited condition characterized by abnormal build up of a protein called amyloid in the body. DefiniGEN hepatocytes present a good model system that mimic the disease phenotype and allow researchers to understand important disease-related mechanisms. This graph shows expression of the TTR pathway in our control wild type hepatocytes as comparable levels to human primary cells.
Alagille Syndrome
Alagille Syndrome is caused by mutations in the JAG1 and NOTCH2 genes. The NOTCH2 and JAG1 gene expression is shown here to be at comparable levels to human primary cells in DefiniGEN’s differentiated hepatocytes, and therefore provide a good isogenic control to our disease model cells.
Hereditary Haemochromatosis
The HFE gene is central in hepatocytes for enabling the cells to effectively metabolize iron, and mutations in this gene can lead to Haemochromatosis. The HFE pathway is shown here by qPCR to be expressed at sufficient levels in DefiniGEN’s differentiated hepatocytes, to provide a good isogenic control to our disease model cells.
Frequently Asked Questions
CRISPR Modeling
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It depends on the complexity of the project but generally we deliver the edited cell line in 8 weeks. If differentiation into a specific cell type is required, additional time will be necessary and will depend on the requested scale and cell type of interest.
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We normally design up to 3 guide RNAs per model using software powered by the latest scoring algorithms. Depending on cutting efficiency (TIDE or ICE analysis), predicted on-target and off-target score we select the best guide for gene editing.
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We work as a research partner offering collaborative services. We offer bi-weekly calls with our clients and interim emails with project reports. Our team is always available to provide technical support which can include experimental design and training on best practices for handling cells. Contact us today >
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To generate a quote, we need your name, the name of your organization, your email address and details on the gene, mutation of interest, and cell type to edit. We also recommend contacting our team to go through project requirements in more detail. Contact us today >
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We perform Sanger sequencing to show successful introduction of frameshift indels in the gene of interest. Additionally, we can evaluate protein expression via Western Blot or perform basic phenotypic screening.