Gene Editing Service - Definigen
Vials in Dry Ice

Gene Editing

DefiniGEN is highly experienced in using CRISPR/Cas9 gene editing technology to manipulate the genome of iPSC to generate disease models

Why choose DefiniGEN?

Our expertise can help design the guide RNA’s (gRNA) in silico for maximum efficiency, then optimize the electroporation of the cell line in question to best establish ideal conditions for the gene edit. Our team then methodically sequence a number of clones to confirm the precise genetic manipulation has been successful.

Edits available

Service Description Applications
Knockout
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
Knock-in
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 casettes integration Introducing large casettes
Tag Reporter Integration of reporters or tags
Scientist Definigen

Gene editing service process

  • Optimal sgRNA design – screening 3 sgRNA
  • Optimal donor design – to maximize the efficiency of knock-ins/knockouts: selecting optimal homology arm length, amount of donor DNA, donor strand orientation and distance between cutting and insertion site
  • Choice of donor template: ssDNA or dsDNA - Choice of donor type is often dictated by the size of the modifications to be introduced: ssDNA donors have been mostly used for applications requiring small edits, dsDNA are used for integration of gene-sized reporters. ssDNA donors are advantageous to achieve a higher specificity of integration, in a direct comparison with equivalent dsDNA sequences.
  • Delivery method: via Electroporation
  • Cell line: wide range of cells
  • Genotyping/screening: clonal (homozygous/heterozygous modification) or pooled

Gene Editing Workflow

Gene-editing of iPSCs allows researchers to create isogenic cell models containing key disease driving mutations to achieve mechanistic understanding, without background genetic variability allowing determination of causative relationships between genotype and phenotype.

Scientist Pipetting

Deliverables

  • Highly characterized/validated edited human cell lines (clonal or pool)
  • Isogenic (parental) controls to remove any phenotypic effects driven by factors intrinsic to the particular mutation of interest