CRISPR Gene Editing Service
Maximize the power of iPSC technology with our CRISPR gene editing service
Why choose DefiniGEN?
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.
Learn about our combined iPSC gene editing and differentiation services here >
We are an approved supplier on scientist.com
Our CRISPR Gene Editing Services
Whether you need simple gene knockouts, knock-ins or more sophisticated genome editing, our scientists can deliver.
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Knock-in Gene Editing ServiceStudy disease-causing mutations
We can introduce point mutations/SNPs or correct disease-causing mutations. We can also insert large cassettes, as well as reporters or tags to facilitate your disease research.Learn more -
Knock-out Gene Editing ServiceInactivate genes to study their function
We can inactivate your target gene by inserting a disruptive indel or by deleting a large section (few bp to > 1kb) of the target gene. We can also apply these methods to knock out multiple genes in the same reaction.Learn more -
IPSC DifferentiationWe've transformed the "difficult" into the "routine" and have successfully differentiated multiple iPSC lines into a range of target cell types that fully recapitulate the mature cell phenotype.Learn More
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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.
Gene Editing Workflow
*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.
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Project initiationWe 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.
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Targeting sgRNA designGene editing success starts with good sgRNA design. We streamline the process by screening sgRNA sequences in silico and selecting the top three sgRNAs for in-vitro screening. -
Donor template designTo maximize the efficiency of knock-ins and knock-outs, we optimize the lengths of the homology arms, the amount of donor DNA, donor strand orientation, and the distance between the CRISPR cleavage site and the mutation site.
We use ssDNA donors when making point mutation or small edits of only a few nucleotides in length, and plasmid donors for larger edits. -
Gene editingOnce your custom reagents are created, we optimize the electroporation conditions to ensure robust delivery of all CRISPR gene editing components.
We screen the edited population at pool level to check cutting efficiency, seed single cells, and sanger sequence individual clones to identify successfully edited cells. -
Product validationWe use a variety of analytical tools, including genotyping, phenotyping, and cell-based assays, to ensure that the cells contain the desired edits and retain pluripotency. -
Delivery of modified iPSCsUpon successful validation, we ship your cryopreserved iPSCs to you, and you can expect high viability post-thaw.
Experience with a range of iPSC edits
| Cell Type | Project Description |
| iPSC-derived hepatocytes |
We have used gene editing of iPSCs to develop optimized hepatocyte models of a range of diseases:
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| iPSC-derived pancreatic cells |
We have used gene editing of iPSCs to systematically introduce known gene variants associated with type 2 diabetes into pancreatic cells, as well as the MODY and neonatal forms of diabetes. |
Frequently Asked Questions
Gene Editing Service
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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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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.
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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.
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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.
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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.
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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.
