Transgenic animals

Generation and analysis of gene modified mice

The BCO Transgenic (BCO-Tg) Core Facility coordinates the Finnish participation in the Infrafrontier ESFRI project and serves as the Finnish node of the European Mouse Mutant Archive (EMMA). www.infrafrontier.eu In addition, it coordinates the activities of the Biocenter Finland Model Organisms infrastructure network & technology platform services www.biocenter.fi, the BF FinGMouse network www.fingmice.org and the NordForsk-funded Nordic Infrastructure for Mouse Models, NorIMM.

Furthermore, the BCO-Tg core facility provides a wide range of other services related to genetically modified (GM) mice:

  1. Gene targeting and culture of embryonic stem (ES) cells
  2. Generation of chimeric mice by ES cell injections
  3. Generation of transgenic mice by DNA microinjection into zygotes
  4. Rederivation of contaminated mouse lines
  5. Cryopreservation of mouse lines
    - Infrafrontier/EMMA services
    - Other cryopreservation services
  6. Recovery of cryopreserved mouse lines
  7. Generation of induced pluripotent stem (iPS) cells
  8. Isolation of ES cells
  9. Lentivirus injections into zygotes
  10. Mouse phenotyping
  11. Service prices 2017

Contact persons:

Coordinator Raija Soininen, Ph.D., Docent
tel. +358-(0)294 486135
e-mail: raija.soininen (at) oulu.fi

Researcher Reetta Vuolteenaho, Ph.D.
tel. +358-(0)294 486133
e-mail: reetta.vuolteenaho (at) oulu.fi 

1. Gene targeting and culture of embryonic stem (ES) cells

ES cell lines available in the core facility are derived from 129S6, 129X1, and C57BL/6N strains, all of them with high germ line efficiency. The customer provides the linearized construct DNA to the core facility and plans and tests the genotyping protocol.

ES cell culture, electroporation, selection, colony picking and freezing, as well as isolation of DNA from cell clones are performed by the Transgenic Core Facility staff. The customer is responsible for the DNA analysis. Clones carrying the targeted alteration are then expanded and frozen for further use.

The success in the targeting is dependent on the gene region and the targeting construct, therefore, a positive result cannot be guaranteed but is in most cases obtained.

The price of one targeting project includes the picking, freezing 24-well plates, and DNA isolation of about 200 colonies. Thawing and expansion of ES cell clones and freezing in vials for long-term storage in liquid nitrogen is charged separately.

Order Form

2. Generation of chimeric mice by ES cell injections

The customer has to have a valid license for generation of a new mutant mouse line before the injections can be started (ELLA).

The core facility prepares the ES cells for injection. If the ES cells were grown outside the core facility, only cells with a clean mycoplasma test result are accepted. The result is confirmed in another test in the core facility.

Injections of the cells into C57BL/6N blastocyst or morula-stage embryos, and transfer into recipient females are performed by the core facility staff. If C57BL/6 ES cells are used, the core facility personnel genotypes the pups to identify the chimeric mice. With other cells, the fur color is the indicator for chimerism.

The price of one project includes the injection of cells of one targeted ES cell clone, transfer of 70-80 embryos into foster mothers and mouse care until weaning.

Chimeric mice are delivered to the customer, or if agreed with the Transgenic core and the Laboratory Animal Center, are mated with wild type mice in the barrier. The customer genotypes the offspring of the chimeras and plans further breeding.

Price for breeding: please contact Raija Soininen.

Order Form

3. DNA microinjections into zygotes

The customer has to have a valid license for generation of a new mutant mouse line before the injections can be started (ELLA).

The customer prepares the construct, plans and tests the genotyping, and supplies the purified DNA. For the purification protocol, click here. Microinjections and transfer of embryos into recipient mothers are performed by the core facility staff.

Mouse strains routinely used for zygote injections are currently FVB and the hybrid C3H/HeN/C57BL/6.

The customer is responsible for the analysis of the pups born, and for planning further breeding.

The price of one project includes the injections of one DNA construct, transfer of 200 embryos into foster mothers and the mouse care until weaning.

Order Form

4. Rederivation of contaminated mouse lines

The core facility and the Laboratory Animal Center have to be contacted well in time for organizing the quarantine space for the mice to be cleaned. Costs for the mouse care in the quarantine are covered by the customer.

If the mouse line to be rederived carries pathogenic viruses, a health screening is performed for the recipient mothers after the pups are weaned, the cost of which will be charged from the customer.

Routinely, the rederivation of a mouse line into specific pathogen free (SPF) condition is performed via embryo transfer so that mutant males provided by the researcher are mated with wild-type females. If the mice carry a complex genotype and wild-type females cannot be used, the customer has to provide 3-5 week-old females carrying the correct genotype.

Cleansing of embryos isolated elsewhere can also be performed.

The price of one project includes the superovulation of 4-6 oocyte donors, transfer of the embryos into foster mothers, and mouse care until weaning.

Order Form

5. Cryopreservation of mouse lines

Archiving of a mouse line includes cryopreservation of sperm and/or embryos in liquid nitrogen.

Infrafrontier/EMMA service: Archiving and distribution of mutant mouse lines

The BCO TG-core facility serves as the Finnish Infrafrontier/EMMA (The European Mouse Mutant Archive) node. Archiving in EMMA is recommended and free of charge. For submission and ordering of EMMA mouse strains, please see www.infrafrontier.eu.

If the archiving is done for other than EMMA purposes, the price of a project includes the cryopreservation of sperm from 2-3 males, and the quality control of the frozen sperm by in vitro fertilization (IVF), followed by either in vitro culture or transfer of 2-cell embryos generated. The customer performs the genotyping.  

Cryopreserved embryos can be generated either by IVF or by mating. For the IVF option, the price of a project includes the cryopreservation of both sperm from 2-3 males and 2-cell embryos (up to 200) generated by IVF and as quality control either in vitro culture or transfer of 2-cell embryos. For cryopreservation of embryos generated by mating, the customer has to provide both males and 3-5 week-old females carrying the correct genotype. The price of one project includes the superovulation of 15 oocyte donors, the isolation and cryopreservation of the embryos and as quality control either in vitro culture or transfer of embryos.

Order Form

6. Recovery of cryopreserved mouse lines

The customer has to have a valid license for breeding of the mouse line to be recovered (ELLA).

Cryopreserved mouse lines are recovered by thawing frozen embryos or sperm. If sperm is the cryopreserved material, IVF is used to generate embryos. Embryos are transferred into recipient females. The customer performs the genotyping of the pups. 

The price of one project includes either the IVF or thawing of embryos, transfer of the embryos into foster mothers, and mouse care until weaning.

Order Form

7. Generation of induced pluripotent stem (iPS) cells

Generation of iPS cells is performed by the retrovirus infection method (Okita et al., 2007). The cells to be induced are either provided by the researcher, or the core facility will isolate and culture cells from mice provided by the customer.

Price: depends on the experiment, please contact Raija Soininen.

8. Isolation of ES cells

Isolation and expansion of ES cells can be performed from requested mouse lines using the 2i method (Nichols et al 2009). The mouse line is provided by the researcher. The core facility will isolate the morulae and isolate and culture the ES cells.

Price: depends on the experiment, please contact Raija Soininen.

9. Lentivirus injections into zygotes

Lentivirus injections are performed upon request.

Price: depends on the experiment, please contact Raija Soininen.

10. Mouse phenotyping

For more information, visit this page.

11. Service prices 2017

Services and prices for customers from University of Oulu, please see here.

Gene targeting in ES cells

3500 €

Thawing and expansion of ES cell clones

825 €

Morula injection

3600 €

Pronuclear injection
hybrid or FVB; DNA or RNA

4300 €

Pronuclear injection
C57BL/6N; DNA or RNA

5200 €

Re-derivation into barrier

1200 €

Recovery of a mouse line from frozen embryos

  600 €

Recovery of a mouse line from frozen sperm

1550 €

Cryopreservation of sperm

715 €

Cryopreservation of sperm
(QC IVF + transfers)

2000 €

Last updated: 15.8.2017