CRISPR Conditional Knockout iPSC Service

Cell lines with a loss-of-function gene mutation is an invaluable tool for studying and understanding function of genes and gene products. But a constitute knockout of an essential gene can affect cell viability and survivability. A conditional knockout cell line could rescue a potential lethality and give better control over the expression of the gene, as well as offer a more physiologically relevant model to study the role of the target gene and provide a target for drug discovery.

Leverage Applied StemCell unique expertise in CRISPR iPSC/ ES Cell Genome Editing Service to generate a conditional knockout iPSC line. We can generate conditional gene knockout in your gene of interest.

• Send us your healthy/ patient iPSC or ESC lines; OR
• Choose from our well-characterized control “master” iPSC lines

Also available, downstream iPSC differentiation and characterization services.

• Optimized CRISPR protocols for high success rate
• Homozygous/ heterozygous conditional knockout clones
• Footprint-free, feeder-free transfection and iPSC culture protocols
• Isogenic control lines for reliable experiments

Workflow includes:

• Cell line evaluation
• gRNA design, CRISPR vector and donor DNA construction, and reagent validation
• Transfection of targeting vectors and optimization
• Screening for single cell clones and clone confirmation
• Cell expansion and cryopreservation

Deliverables:

• Two (2) clones with confirmed modification(s), with two (2) vials of each clone at 1 x 10^6 cells/vial.
• Project Milestones Reports, as well as a Final Report containing details of targeting design, experimental, and genotyping results.

Timeline: 3-4 months

Applications:

• Physiologically relevant disease models for hard-to-model diseases (Ex. ALS, muscular dystrophy, Parkinson's disease, Alzheimer's)
• Differentiate to study mutations in different tissue lineages with an isogenic panel of cell line models
• Ideal for target drug discovery, drug and toxicity screening