Mouse Model Generation by the Biocenter at the University of Oulu

Transgenic and Tissue Phenotyping Core Facility, Biocenter Oulu, University of Oulu (Finland)

Biocenter Oulu research infrastructure operates at the University of Oulu, Finland, and serves academic and non-academic customers in research and R&D projects. The Transgenic and Tissue Phenotyping (TTP) Core Facility provides a wide range of services for planning, generation, archiving, recovery, and re-derivation of gene-modified (GM) mouse lines as well as for tissue phenotyping: histology, digital pathology, and image analysis. TTP Core Facility serves as the Finnish European Mouse Mutant Archive (EMMA) node in INFRAFRONTIER ( www.infrafrontier.eu/) and coordinates the national activities of the Biocenter Finland FinGMice network. As an EMMA service, TTP core provides archiving and distribution of GM mouse lines without costs to provider scientists

Photo credit: Mikko Törmänen

Mouse Model Generation Services offered by the Transgenic and Tissue Phenotyping Core Facility:

1. CRISPR/Cas9 editing (for academic customers)

Service Description:

CRISPR-Cas9 technology enables the modification of genome to produce precise and unique knock-out and knock-in GM mouse models. Possible allele types that can be generated are indels, exon deletions (< 10kb), point mutations and insertions.

Included in the Service:

design of the guides (crRNA+tracrRNA),
design of ssODN or lsODN DNA templates,
prediction of off-targets,
electroporation or microinjection of guide RNA/Cas9 protein mix and template DNA (knock-in only) into C57BL/6N or C57BL/6J zygotes,
transfer of up to 200 embryos into recipient mice,
mouse care in a specific pathogen free environment,
genotyping of founders and Sanger sequencing of the target locus,
breeding of founders (2-4 per line),
genotyping of F1 mice and Sanger sequencing of the target locus, and
organising shipment of generated mice to the receiving institute (shipping costs covered by the customer)

The customer is responsible for providing synthetic guide RNA, DNA template and Cas9 protein.

Additional Support with separate pricing offered:

sequencing of the off-target sites on the same chromosome, and
breeding of F1 animals (2-4) and genotyping of F2 generation.

2.a. Gene targeting in embryonic stem cells

Service Description:

Mouse embryonic stem (ES) cells are cultured in pluripotent stage and mutations targeted to a specific locus to produce knock-out and knock-in GM mouse models. ES cell lines derived from 129S6 and C57BL/6N strains are available at the core.

Included in the Service:

ES cell culture, electroporation, selection, colony picking, freezing and DNA extraction from up to 200 colonies, and
organising shipment of genotyping samples

The customer is responsible for designing and providing linearized construct DNA as well as genotyping of ES cell clones.

Additional Support with separate pricing offered:

genotyping of ES cell clones,
expansion of correctly targeted ES cell clones and freezing in vials for long-term storage, and
organising shipment of ES cells.

2.b. ES cell injection into morula-stage embryos

Service Description:

Correctly targeted ES cells are microinjected into morula-stage mouse embryos to produce chimeric mice. When ES cells differentiate into germ cells, targeted mutation becomes inheritable resulting in the establishment of a knock-out and knock-in GM mouse model.

Included in the Service:

ES cell culture and microinjection of one targeted ES cell clone into C57BL/6N or C57BL/6J morulae,
transfer of 70-80 embryos into recipient mice,
mouse care in a specific pathogen free environment, and
organising shipment of generated mice to the receiving institute (shipping costs covered by the customer)

Additional Support with separate pricing offered:

mycoplasma testing of imported ES cells (only mycoplasma-free cells are accepted),
breeding of chimeric mice, and
genotyping.

3. DNA injection to generate transgenic models

Service Description:

Typically, a transgene construct is composed of a promoter, coding region and 3’ untranslated sequences. In transgenic GM mouse models, the foreign DNA introduced is integrated into the genome in a random manner and often as multiple copies.

Included in the Service:

microinjection of a transgene construct into C57BL/6N or C57BL/6J zygotes,
transfer of up to 200 embryos into recipient mice,
mouse care in a specific pathogen free environment, and
organising shipment of generated mice to the receiving institute (shipping costs covered by the customer)

The customer is responsible for designing, preparing and purification of the DNA construct.

Additional Support with separate pricing offered:

breeding of founder mice, and
genotyping

Relevant Publications

Ravindran A, Holappa L, Niskanen H, et al. Translatome profiling reveals Itih4 as a novel smooth muscle cell-specific gene in atherosclerosis. Cardiovasc Res. 2024; 120(8):869-882. doi: 10.1093/cvr/cvae028.
Brouillard P, Murtomäki A, Leppänen VM, et al. Loss-of-function mutations of the TIE1 receptor tyrosine kinase cause late-onset primary lymphedema. J Clin Invest. 2024; 134(14):e173586. doi: 10.1172/JCI173586.
Kapiainen E, Elamaa H, Miinalainen I, et al. Cooperation of Angiopoietin-2 and Angiopoietin-4 in Schlemm’s Canal Maintenance. Invest Ophthalmol Vis Sci. 2022; 63(11):1. doi: 10.1167/iovs.63.11.1
Hiltunen AE, Kangas SM, Ohlmeier S, et al. Variant in NHLRC2 leads to increased hnRNP C2 in developing neurons and the hippocampus of a mouse model of FINCA disease. Mol Med. 2020; 26(1):123. doi: 10.1186/s10020-020-00245-4

How to apply to the INFRAFRONTIER Mouse Model Generation service?

Please use INFRAFRONTIER Service Request Form or Pbagnpg hf if questions.