Generation of mutant mouse lines

Knock-out models

Knock-in models

Transgenic models


Knock-out mouse model: CRISPR/Cas9 editing in zygotes

Microinjection/electroporation of gRNA/Cas9 protein mix into zygotes and embryo transfer are performed by the core.

Mouse backgrounds used are C57BL/6NCrl and C3HB6F1/N hybrid.

The customer is responsible for:

  • having a valid license for generation of a new mutant mouse line (ELLA)
  • providing the synthetic guide RNA and Cas9 protein
  • the genotyping of the pups born
  • planning further breeding

The service includes the injections/electroporation and transfer of up to 200 embryos into foster mothers, and mouse care until weaning.

Knock-out mouse model: Gene targeting in embryonic stem cells

Culturing of ES cells and the electroporation of DNA construct are performed by the core.

ES cell lines available in the core facility are derived from 129S6, 129X1, and C57BL/6N strains.

The customer is responsible for:

  • having a valid license for generation of a new mutant mouse line (ELLA)
  • designing and providing the linearized construct DNA. For DNA preparation: click here
  • the genotyping of the ES cells

Service includes culturing of ES cells, electroporation, selection, colony picking, freezing and extraction of DNA from up to 200 colonies. Clones carrying the targeted alteration are then expanded and frozen for further use.

Thawing and expansion of ES cell clones and freezing in vials for long-term storage in liquid nitrogen is charged separately.

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.

Knock-out mouse model: ES cell injections to morula-stage embryos

Culturing of ES cells, injections of the cells into C57BL/6NCrl morulas and embryos transfers are performed by the core.

The customer is responsible for:

  • having a valid license for generation of a new mutant mouse line (ELLA)
  • If the ES cells were imported to the researcher/core facility, only cells with a clean mycoplasma test result are accepted
  • the genotyping of the pups. For genotyping: click here
  • for planning further breeding

Service includes culturing (mycoplasma testing of imported ES cells) and injection of one targeted ES cell clone, transfer of 70-80 embryos into foster mothers and mouse care until weaning. If C57BL/6N 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.

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.

Knock-in mouse model: CRISPR/Cas9 editing in zygotes

Microinjection/electroporation of gRNA/template DNA/Cas9 protein mix and embryo transfer are performed by the core.

Mouse backgrounds used are C57BL/6NCrl and C3HB6F1/N hybrid.

The customer is responsible for:

  • having a valid license for generation of a new mutant mouse line (ELLA)
  • providing the synthetic guide RNA, template DNA and Cas9 protein
  • the genotyping of the pups born
  • for planning further breeding

The service includes the injections/electroporation and transfer of up to 200 embryos into foster mothers, and mouse care until weaning.

Knock-in mouse model: Gene targeting in embryonic stem cells

Culturing of ES cells and the electroporation of DNA construct are performed by the core.

ES cell lines available in the core facility are derived from 129S6, 129X1, and C57BL/6N strains.

The customer is responsible for:

  • having a valid license for generation of a new mutant mouse line (ELLA)
  • designing and providing the linearized construct DNA. For DNA preparation: click here
  • the genotyping of the ES cells

Service includes culturing of ES cells, electroporation, selection, colony picking, freezing and extraction of DNA from up to 200 colonies. Clones carrying the targeted alteration are then expanded and frozen for further use.

Thawing and expansion of ES cell clones and freezing in vials for long-term storage in liquid nitrogen is charged separately.

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.

Knock-in mouse model: ES cell injections to morula-stage embryos

Culturing of ES cells, injections of the cells into C57BL/6NCrl morulas and embryos transfers are performed by the core.

The customer is responsible for:

  • having a valid license for generation of a new mutant mouse line (ELLA)
  • If the ES cells were imported to the researcher/core facility, only cells with a clean mycoplasma test result are accepted
  • the genotyping of the pups. For genotyping: click here
  • for planning further breeding

Service includes culturing (mycoplasma testing of imported ES cells) and injection of one targeted ES cell clone, transfer of 70-80 embryos into foster mothers and mouse care until weaning. If C57BL/6N 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.

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.

Transgenic models

Microinjection of the transgene construct into zygotes and embryo transfers are performed by the core.

Mouse backgrounds used are C57BL/6NCrl and C3HB6F1/N hybrid.

The customer is responsible for:

  • having a valid license for generation of a new mutant mouse line (ELLA)
  • designing, preparing and purification of the DNA construct. For the purification protocol, click here.
  • the genotyping of the pups born
  • for planning further breeding

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

Last updated: 30.3.2020