Stem Cell Facility - Ahus

• Project planning and consultation:

Experimental design (in different cell types and species), Designing of guide RNAs, sequence annotation and virtual editing maps (SNAPgene, Benchling etc), primer designing, and editing efficiency of guide RNAs (ICE analysis, TIDE analysis etc). Cooperation during manuscript writing. Project planning comes along with any of the services listed below.

• Knock Out editing: Designing of guide RNAs (CRISPOR, CHOP CHOP, Benchling etc), electroporation (Neon and Amaxa based), editing efficiency by Sanger sequencing and analysis tools (ICE analysis, TIDE analysis etc), single cell seeding using IOTA (isohub), pure clone selection, re-confirmation of the knockout (Sanger sequencing and RFLP based), master bank generation, pure clones scale-up and freezing.

• Knock-in generation: Designing of guide RNAs, designing of ssODN donor/plasmid donor for knock-in HDR (Manual, Benchling etc), electroporation (sgRNA+Cas9+ssODN/plasmid), editing efficiency by Sanger sequencing analysis, single cell seeding, pure clone selection after single cell seeding, re-confirmation of the knock-in (Sanger sequencing and RFLP based), master bank generation, pure clones scale-up and freezing. 

• Point mutation (SNP) correction/insertion: Designing of guide RNAs, designing of ssODN donor (correction template), electroporation (sgRNA+Cas9+ssODN/plasmid), editing efficiency by Sanger sequencing, single cell seeding, pure clone selection using single cell seeding, re-confirmation of the correction/insertion (Sanger sequencing and RFLP based), master bank generation. 

• Reporter cell line generation: Designing of guide RNAs, designing of donor template for reporter (GFP, RFP reporter etc), electroporation (sgRNA+Cas9+ssODN/plasmid), editing efficiency by Sanger sequencing, single cell seeding, pure clone selection using single cell seeding, re-confirmation of the correction/insertion (Sanger sequencing and RFLP based), master bank generation.
• Base editing in iPSCs- Base editing represents a cutting-edge genome editing technique that combines CRISPR system components with additional enzymes to precisely introduce point mutations into cellular DNA or RNA, bypassing the need for creating double-stranded DNA breaks (DSBs): coming soon
• Prime editing in iPSCs- While base editing excels at introducing specific nucleotide changes like transition and transversion mutations within the targeted region, prime editing offers a broader capability by employing a "search-and-replace" mechanism to create precise insertions, deletions, and all 12 types of point mutations- coming soon

Prices

Optional services:

• Extra cell stocks (24 or 48 extra vials)
• Additional edited or unedited clones (3000 NOK/extra clone, 5 vials)

After completion of the project, we will provide at least five vials of two edited clones (0.5 million cells per vial).

IOTA single cell dispenser

   

Isohub system for single cell cloning

Isohub microscopes with various LEDs for fluorescent microscopy

Neon Electroporation system