Molecular Services

In association with our mouse Embryonic Stem Cell and Transgenic Core Services facilities, the Molecular Services Core offers designing, construction and preparation of vectors for the generation of genetically engineered mice.  This includes CRISPR KI/KO/CKO in mouse lines.  Contact us to discuss your project needs.

The Molecular Biology Core also offers CRISPR gene knock-out services for cell lines.

Our services include but are not limited to:

Genotyping (including genomic DNA preparation)

• PCR screening, genotyping, genotyping protocol
• PCR genotyping program trouble shooting and redesigning
• Long Range-PCR (6-7kb)

Designing, Construction and Preparation of Transgene Vector

• Plasmid
• BAC

Designing, Construction and Preparation of Gene Targeting Vector

• Conventional knock-out
• Conditional knock-out
• Conditional restoration
• Knock-in (e.g., point mutation, humanization, reporter, ROSA26)
• Conditional expression of transgene from ROSA26 locus

Designing, Construction and Preparation of Artificial Nucleases (ZFN, TALENs, CRISPRs) for Microinjection into Mouse Embryos

• TALENS, ZFN (ROSA)
• CRISPR/Cas9
• Designing and construction of donor DNA (linear ds & ss DNA, plasmid, BAC and minicircle)
• CRISPR generated Gene knock-out in mouse embryos
• Designed modification of short genomic sequence in mouse embryos

Custom Somatic Cell CRISPR Gene Knock-out ~$5,000.00 ~3 months timeline
Since 2016 we have completed more than 43 CRISPR gene knockout projects in 34 different cell lines targeting 25 different genes for researchers located at the University of Calgary and University of Alberta.

• Guide RNA design, construct and plasmid preparation
• Transfection with guide RNA, Cas9
• Use of selection markers such as puromycin and GFP to enrich for transfected cells
• CRISPR activity analysis: SURVEYOR mutation detection assay
• Single clone generation: sorting cells plus expansion
• Gene knock out confirmation by Western blot analysis
• Gene knock out confirmation by DNA sequencing
• Final deliverable: gene knock out cell line and a complete report

For researchers who may not have the necessary molecular biology background, the opening of our Molecular Biology Core Facility at the CGE makes the generation of novel mouse models accessible to everyone.

Contact us:

Robyn Flynn  -  molbiol@ucalgary.ca (genotyping, mouse related molecular services)

Yaping Yu  -  yuy@ucalgary.ca (custom somatic cell CRISPR knock-out services)

General questions at transgen@ucalgary.ca