Delivering phenotypically relevant disease models


Expand your insight into cellular physiology and gene function with knock-in cell lines



When you're ready to study disease causing mutations in a clinical context or protein function in a native cell biology, DefiniGEN can advance your studies through efficient creation of an appropriate knock-in cell line. 


Combining years of experience handling iPSCs with an understanding of how iPS cells respond to CRISPR-based genome editing, we deliver high success rates and a collaborative customer experience.



Our optimized protocols and standardized workflows ensure fast, efficient project completion.





Our deep expertise handling challenging iPSCs easily translates into successful CRISPR editing of iPSCs.





We validate successful knock-in cell line generation through a variety of assays including genotyping, phenotyping, and cell marker analysis.


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Knock-in workflow


Because every cell line responds differently to CRISPR gene-editing, we start each project with an evaluation of your iPS cell line's response to editing. alternatively, you can accelerate turnaround times and maximize success by starting with one of our in-house cell lines that do not require as extensive an evaluation. 


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.



How it works

  • We design up to 3 single guide RNAs (sgRNAs) per target gene using software powered by the latest scoring algorithms.

  • Depending on their cutting efficiency and predicted on-target and off-target scores, we select the best sgRNA for gene editing.

  • Purified, high-fidelity CAS9 protein is delivered into your chosen cells together with the optimal sgRNA in the form of ribonucleoprotein (RNP) complexes.

  • Once CRISPR editing is complete, we perform an array of standard QC and validation tests on the knock-in cells, and several additional services are also available depending on your project needs. 

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QC Testing and Additional Services

We offer an array of validation services to fully characterize your knock-in cell lines.



Service Description

Morphology, Sterility, and Pluripotency

We confirm that your cells have the expected morphology, express pluripotency mRNAs, and are sterility tested.

Mycoplasma Testing

We confirm that your knock-in cells are free from mycoplasma contamination

qPCR Screening

Validation of your knock-in cells to confirm they carry the target deletion/indel on the coding sequence at the mRNA level

Additional Services

Depending on your project needs, we can perform any of the following additional validation assays on your knock-in cell lines


RNAseq provides insight regarding the impact of CRISPR gene knock-in on a global transcriptomic scale.

iPSC Differentiation

We can save you time and costs by differentiating your CRISPR-edited iPSC lines for you using our highly efficient differentiation platform.

Off-target analysis

Additional off-target screening is available using in silico prediction, targeted sequencing, exome sequencing, or whole-genome sequencing.


Technical Support From Start to Finish

We pride ourselves on working with each client as a collaborative research partner!


What you will receive:

  • Bi-weekly calls and interim emails to update you on the project progress.
  • Highly characterized/validated CRISPR-edited human cell lines (clonal or pool).
  • Final report containing details of the sgRNA design and the results of all QC and validation assays.
  • Isogenic (parental) controls to determine any phenotypic effects driven by factors intrinsic to the gene knock-in.

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Frequently asked questions


How long does the entire gene-editing process take?

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.

How do you confirm that the cells have the correct modification?

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.

Which gene editing method do you use?

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.

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

Inactivate 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.

We'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.

Advance your scientific discoveries with our tailored disease modeling products and services.