Gene Editing Services
Unlock advanced research possibilities with our CRISPR/Cas9-based gene editing knock-in and knock-out solutions
Accelerate your projects with our expert-driven genome editing services
From permanent or conditional knock-outs to point mutations and targeted insertions, we deliver precision modifications aligned with your research needs.
Combining years of experience handling induced pluripotent stem cells (iPSCs) with an expert understanding of how iPSCs respond to CRISPR-based genome editing, we deliver high success rates and a collaborative customer experience.
Advantages of working with DefiniGEN
Rapid turnaround time
Our optimized protocols and standardized workflows ensure fast, efficient project completion.
High success rate
Our deep expertise handling challenging iPSCs easily translates into successful CRISPR editing of iPSCs.
Functional Validation
We validate successful knock-in cell line generation through a variety of assays including genotyping, phenotyping, and cell marker analysis.
Scientist.com approved
We are an approved supplier on scientist.com. Our services have been validated and approved through scientist.com
Our Gene Editing workflow
Every cell line responds differently to CRISPR gene-editing, which is why we start each project with an evaluation of your iPSC line's response to editing.
Additionally, you can accelerate turnaround times and maximize success by selecting one of our highly characterized in-house cell lines.
Cell line evaluation
If you choose one of our cell lines for editing, we will sequence the locus of interest, alternatively if you wish to provide your own cells for editing we will assess their proliferation rate.
gRNA design, screening, and construction
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.
Transfection and optimization
Purified, high-fidelity CAS9 protein is delivered into your chosen cells together with the optimal sgRNA in the form of ribonucleoprotein (RNP) complexes.
Screening for single positive clones
We screen edited cells via sanger sequencing and perform alignment analysis to reference the sequence using Benchling.
Cell expansion and validation
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.
Delivery
Delivery of edited cells will take approximately 4 weeks.
Quality Control 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 |
|
RNA-Seq |
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! 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 Quality Control and validation assays.
- Isogenic (parental) controls to determine any phenotypic effects driven by factors intrinsic to the gene knock-in.
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.
