GENERATION OF HUMAN IPSC
The platform proposes the reprogramming of somatic cells (human fibroblasts or blood cells) into iPSc using Sendai vectors and the full molecular and functional characterization of three of the iPSc clones generated.
The characterization comprises:
- Verification of pluripotency and clearance of Sendai virus by Immunofluorescence and RT-Q-PCR (Scorecard analyses)
- Verification of chromosomal integrity by iCS-digitalTM Pluri and SNP Array (optional)
- Quality test: viability and mycoplasma control
GENETIC ENGINEERING OF HUMAN IPSC
The platform uses the CRISPR/Cas9 technology to modify iPSc clones (NHEJ or HR).
The service comprises:
- Design of CRISPR tools:
- ssODN (point mutation correction/insertion for example)
- For longer insertion (tagging project for example), collaboration with the vectorology platform for the design/construction and production of the HR plasmid
- iPSc line editing and clone analyses by PCR and sequencing
- Expansion of 1 to 3 modified clones
- Verification of off-target sites if necessary and chromosomal integrity by iCS-digitalTM Pluri