The CRISPR-Cas9 system is able to achieve highly specific and precise targeting for the genetic manipulation of cells and cell lines, transforming the field of genome engineering and presenting a wealth of new possibilities for R&D.
DefiniGEN have optimized conditions for CRISPR Cas-9 genome editing on multiple patient-derived iPSC’s, and we have built up considerable experience in using the system for genomic manipulation in a range of other cell lines.
We can offer our clients an integrated end-to-end package including sgDNA design, transfection, optimisation, and clone picking/sequencing to generate a library of gene edits for client selection. From a basic frameshift knock-out mutation to complex knock-in genetic changes, our dedicated scientific team will work collaboratively with you to design the right tools to accelerate your research programs.
|Indel for gene disruption||Gene inactivation by introducing insertion or deletion||Study gene function via gene inactivation|
|Large deletions||Deletion from few base pairs to kilobase in the desired gene|
|Multiple deletions||Editing with multiple guides in a single reaction|
|Point mutations/SNPs||Introducing point mutation or correcting disease-causing mutation||Study disease causing mutations in a clinical context and protein function in a native cell biology setting|
|Large cassettes integration||Introducing large cassettes|
|Tag Reporter||Integration of reporters or tags|
DefiniGEN's iPSC differentiation platform coupled with CRISPR gene-editing allows the phenotypic modelling of a wide range of metabolic diseases.
Our CRISPR services gives us the ability to repair causative mutations to patient-derived lines, or introduce alleles into a healthy parental line. The differentiated disease model human cells are a powerful tool to support preclinical research.
|Acute intermittent porphyria|
|Genetic cholestasis (PFIC, TGP2, and Alagille syndrome)||Familial hypercholesterolemia|
|Wilson’s disease||Organic acidurias (except MSUD)|
|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 (except ASL)||Citrullinemia|
|Crigler-Najjar syndrome||Familial amyloid polyneuropathy|
|Primary hyperoxaluria type 1||Atypical haemolytic uremic syndrome 1|
|Maple syrup urine disease (MSUD)|
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 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 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 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 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.
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.