Our service focuses on introducing INDELs (frame-shift mutations) to generate gene knock-out lines (KO), each targeting a single gene in either hPSCs or immortalized/cancer cell lines.
Characterization of each engineered cell line includes:
- Cell identity confirmation using STR analysis
- Confirmation of desired editing via Sanger sequencing.
- Karyotyping (Q-banding) and identification of Copy Number Variations (CNVs) at high resolution
- Master bank post-thaw viability and mycoplasma testing.
- Optional evaluation of undifferentiated stem cell markers and pluripotency markers upon 3-germ layer differentiation assay.
We accept cell lines in a cryopreserved state, with a minimum of 1 x 10^6 cells per cryovial.
We generate up to 3 homozygous knockout (KO) clones. Heterozygous KO clones can be provided upon agreement with the applicant. If technically feasible, clones will be analyzed for the occurrence of allelic dropout.
The service typically requires 2 to 3 months for completion and includes the delivery of 1-3 clones, each provided in 10 cryopreserved vials.