Induced pluripotent stem cell (iPSC) technology has enabled the generation of cell-based models that recapitulate human disease. But there are times when you can't find the right iPSC line, or you already have a disease-state iPSC line and need healthy isogenic cells. DefiniGEN can help.
Combining our deep expertise in handling iPSCs with CRISPR genome editing technology, our Gene Editing Service team can design and create iPSCs that are edited to meet your project's needs.
|Cell Type||Project Description|
We have used gene editing of iPSCs to develop optimized hepatocyte models of a range of diseases:
|iPSC-derived pancreatic cells|
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 >
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 >
The entire gene editing process takes between 8-10 weeks depending on the complexity of the project. We will advise you of the lead time when we generate your quote.
We use Sanger sequencing to verify that the correct modifications have been made. Upon request, we can perform whole exome sequencing to scan for off-target edits.
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.
CRISPR is a powerful technology that revolutionised the genome engineering field since its discovery in 2013, and quickly became the method of choice for various genome-targeting purposes. CRISPR gene editing technology is better than other gene editing approaches such as meganucleases, transcription activator-like effector nucleases (TALENs) and zinc finger nucleases (ZFNs), due to the technology's robustness, cost-effectiveness, high specificity, efficiency, ability to edit difficult regions, multiplexing capability and simplicity. In summary, CRISPR allows us to efficiently manipulate genes in versatile ways.
We most commonly use a CRISPR RNP-based gene editing approach. To guarantee rapid and highly efficient gene inactivation (gene knockout), purified CAS9 protein is delivered into cells along with sgRNA (ribonucleoprotein (RNP) complex) via electroporation or transfection.
For knock-in experiments where the goal is either to introduce a point mutation or to insert a reporter/tag, we co-deliver donor repair template (ssODN or dsDNA, respectively) and RNP into the cells to facilitate the double-strand break (DSB)-mediated homology directed repair (HDR). Although we favour the Cas9 RNP delivery approach, we also offer plasmid or viral-based delivery of CRISPR components into the cells.