APN Mouse Model Services
Access to the following Mouse Model services are available
- Overview
- ES Cells to Mouse
- Recessive ENU
- Dominant ENU
- Pathology
- Strain Repository
Overview
The Australian Phenomics Network provides researchers with access to expertise and infrastructure for the creation, characterisation and archiving of mouse models for the study of human and animal disease.
ES Cell to Mouse models
For a brief explanation of ES Cell to Mouse models, click here
For information on Mail-order Modified Mice, see the tab above
ENU models
For a brief explanation of ENU Mouse models, click here
Cryopreservation
Details of the cryopreservation service are shown under the Strain Repository tab above.
Pathology
Details of the pathology service are shown under the Pathology tab above.
ES Cell to Mouse Model
A global initiative is taking place with the aim to systematically inactivate every mouse gene and generate conditional gene-target and/or gene-trap mouse models for all 25,000+ genes in the mouse genome. This will enable scientists to subsequently characterize every gene function. Such information will provide invaluable insights into human gene function with wide-ranging clinical implications, including better understanding of diseases and discovering gene targets for therapeutic agents.
An Australian researcher can simply look up the international ES cell databases (listed in “Useful Links” on the ES Cell to Mouse tab), choose the genetically modified (GM) ES cell line(s) of interest, lodge a request with us and within months we can have heterozygote mice delivered to their laboratory.
The Technology
Gene-target and gene-trap vectors are designed to specifically insert into the mouse genome and disrupt gene function; usually resulting in the production of either truncated or no protein from the gene in which it has inserted.
Mouse ES cells are electroporated with the gene-target or gene-trap vectors and the resultant GM ES cell line is then stored in an ES cell library. Once these ES cell lines have been established, they can be microinjected into mouse blastocysts and implanted into pseudopregnant female mice. Resultant pups tested for high level chimerism are bred to homozygosity, at which point the phenotype of the genetically modified mouse can be observed and characterised.
The Service
Monash APN has set up a service whereby all Australian researchers can access the GM ES cell line libraries and have us generate and deliver to them mice that are heterozygote for their desired gene-trap mutation at a heavily subsidized price.
Advantages
The advantages of using this service include the following:
- a time-efficient option for obtaining a GM mouse-model for your gene of interest
- a cost-effective alternative to traditional knock-out strategies
- a complimentary model to compare with full gene “knock-outs” and ENU mutant mice
For further details of this service, please see the ES Cell to Mouse tab.
ENU Models
ENU mutagenesis provides a phenotype driven approach for obtaining mouse models of human conditions. The APN can provide the expertise, scale and resources upon which researchers can significantly expand the scope of their research endeavours.
The Technology
The basis of the ENU mutagenesis approach lies in the chemical mutagen ENU (ethyl-nitrosourea), which incorporates point mutations on a genome wide scale in premeiotic spermatogonial cells. This allows for the production of a number of genetically unique offspring from a single ENU-mutagenised male.
ENU Mutagenesis can be carried out on an inbred strain or on a sensitised background, such as an established transgenic/knockout line.
These libraries of ENU mutagenised mice are then subjected to a robust phenotypic screen. The unique approach provided by the APN enables researchers to custom-design their phenotypic screen, allowing for the isolation of phenovariants that show abnormalities directly related to the researcher’s specific pathway of interest.
Once heritability of the phenotype is confirmed, the chromosomal location of the causative mutation is identified via SNP-based genome scans against a mapping strain. Exonic sequencing then rapidly identifies the SNP of interest.
Advantages
One benefit of the ENU mutagenesis approach is the unbiased identification of novel alleles. Upon initiating the screening process, no assumptions are made as to the physiological role of a gene or its products.
Other advantages offered by this approach are numerous, including:
- Revealing domain specific effects of the gene product,
- Providing a more accurate representation of human conditions through point mutations rather than entire gene knockouts,
- Development of pre-clinical animal models of human disease,
- Revealing previously unknown genetic components and mechanisms,
- Allowing the researcher to focus on their disease or pathway of interest.
Recessive or Dominant
This approach can be applied for discovering mutations that are inherited in both a dominant and recessive manner.
The phenotypic effect of an allele inherited in a recessive manner can only be visualised when the allele has been brought to homozygosity. Details of the recessive ENU model can be found on the Recessive ENU tab.
The phenotypic effect of an allele inherited in a dominant manner can be visualised in the heterozygote state at G1. Details of the dominant ENU model can be found on the Dominant ENU tab.
ES Cells to Mouse Models
The Embryonic Stem (ES) Cell to Mouse service enables Australian researchers to look up the international ES cell databases, choose the genetically modified (GM) ES cell line(s) of interest, then lodge a request, and within months have heterozygote mice delivered to their laboratory.
The service is available through the latest gene-trap and gene-targeting technology: vectors are designed to specifically insert into the mouse genome and disrupt gene function; usually resulting in the production of either truncated or no protein from the gene in which it has inserted.
Mouse ES cells are electroporated with the gene-target or gene-trap vectors and the resultant GM ES cell line is stored in an ES cell library. Once these ES cell lines have been established, they can be microinjected into mouse blastocysts and implanted into pseudopregnant female mice. Resultant chimera mice are then bred to obtain germline transmission of the mutation. Further breeding to homozygosity allows the phenotype of the genetically modified mouse to be observed and characterised.
This service provides ready access to the global initiative to systematically inactivate every mouse gene and generate conditional gene-target and/or gene-trap mouse models for all 25,000+ genes in the mouse genome. Such information will provide invaluable insights into human gene function with clinical implications, including better understanding of diseases and discovering gene targets for therapeutic agents.
The advantages of the service are:
- a time efficient option for obtaining a GM mouse-model for your gene of interest,
- cost-effective alternative to traditional knock-out strategies, and
- a complimentary model to compare with traditional full gene “knock-outs” and ENU mutant mice
The APN ES Cell to Mouse service operates from the Monash University, Melbourne and includes capabilities in molecular biology, tissue culture, embryology and animal husbandry. The scientists in the facility have several years experience in producing genetically modified mice, providing excellent expertise for this operation.
The Australian Phenomics Network provides a number of cutting edge services available to all academic researchers across Australia. One such service is the ES to mouse service.
Cost Structure
Node Involved
Monash Institute of Medical Research at Monash University
Useful Links
- www.eucomm.org
- www.genetrap.org
- www.komp.org
- www.mmrrc.org
- www.sanger.ac.uk/PostGenomics/genetrap
- http://norcomm.phenogenomics.ca/index.htm
Contact:
T: +61 3 9594 7448
Dr Debbie Bianco
F: +61 3 9594 7114
Monash Node Manager
Monash Institute of Medical Research
Monash University
27-31 Wright Street
CLAYTON VIC 3168
E: deborah.bianco@med.monash.edu.au
Recessive ENU Models
The Recessive ENU service provides researchers with ENU gene variant mouse strains that are characterised by recessive inheritance for their phenotypic trait.
ENU mutagenesis is a phenotype driven approach for obtaining mouse models of human conditions. The strength of the ENU mutagenesis approach lies in the chemical mutagen ENU (ethylnitrosourea), which incorporates point mutations on a genome wide scale in premeiotic spermatogonial cells. This allows for the ongoing production of a number of genetically unique offspring from a single ENU-mutagenised male used to create a library of novel gene function variants.
Most of the variant mice will contain point mutations, meaning that they are more reflective of the mutations that naturally occur in the human genome. ENU Mutagenesis can be conducted on an inbred strain or on a sensitized background, such as an established transgenic/knockout line.
These libraries of ENU mutagenised mice are then phenotypically screened for phenotype. This approach enables researchers to custom-design their phenotypic screen, allowing for the isolation of pheno-variants that display characteristics directly related to the researcher’s specific pathway of interest. Pedigrees that do not produce a phenotype of interest in G3 are abandoned, while those that do are carried forward to establish heritability.
This service is accessible through the Australian Phenomics Facility (APF), Canberra, and the Centenary Institute of Cancer Medicine and Cell Biology (The Centenary Institute), Sydney.
The Recessive ENU service provides researcher with access to other NCRIS funded ENU-recessive variants and identified genetic variants of relevance to various areas of study. Through the Centenary Institute, researchers have access to mouse models of relevance to the study of infection and immunity, and the APF covers a broad range of variants. The APN service supplements researchers by providing access to phenotyping equipment, project design and project management services.
Funding from the federal government through the National Collaborative
Research Infrastructure Strategy (NCRIS) will allow researchers to access the APN Recessive ENU derived mice and the related services at a reduced cost.
Nodes Involved
The Australian Phenomics Facility at The Australian National University
The Centenary Institute of Cancer Medicine and Cell Biology
Contacts:
Australian Phenomics Facility
T: +61 2 6125 9117
Dr Michael Dobbie
F: +61 2 6125 1381
APF Scientific Programs Manager
Australian Phenomics Facility
Hugh Ennor Building
Garran Road
ACTON ACT 0200
T: +61 2 6125 9329
Dr Teresa Morgan
F: +61 2 6125 1381
APF Scientific Programs Manager
Australian Phenomics Facility
Hugh Ennor Building
Garran Road
ACTON ACT 0200
Centenary Institute
T: +61 2 9515 5210
Professor Warwick Britton
F: +61 2 9351 3968
Head of Mycobacterial Research Program
Centenary Institute of Cancer Medicine and Cell Biology
Locked Bag No.6
NEWTON NSW 2042
ENU Recessive Screens
As depicted in Figure 1, the first generation (G1) of mice produced from an ENU mutagenised male only carry one copy of any given mutation. While a screen for dominant-acting mutations can be carried out at this stage, a recessive mutation would not exhibit its phenotypic characteristics at G1.
Each recessive pedigree begins with the pairing of two unrelated G1s for the production of G2 offspring, 50% of which are heterozygous for the causative mutation.
By intercrossing G2 siblings, the mutation is brought to homozygosity in 25% of G3 siblings. It is at this stage that the first screen takes place.
Pedigrees that do not produce a phenotype of interest in G3 are abandoned, while those that do are carried forward to establish heritability.
Heritability is confirmed by the emergence of the phenotype of interest in subsequent generations. Once heritability is established, the strain is fixed through several generations of breeding, thus providing an ongoing supply of phenovariants for further characterisation.
Mapping of the causative mutation forms the next stage of the process. Consecutive stages of breeding to a mapping strain introduce recombination events, narrowing the region in which the mutation is located. Discovery of the mutation location forms the final stage of the mapping process.
Figure 1: The strategy for production of ENU mutagenised pedigrees for a recessive screen
Dominant ENU Models
The Dominant ENU service provides researchers with access to ENU gene variant mouse strains that are characterised by dominant inheritance for their phenotypic trait.
ENU mutagenesis is a phenotype driven approach for obtaining mouse models of human conditions. The basis of the ENU mutagenesis approach lies in the chemical mutagen ENU (ethylnitrosourea), which incorporates point mutations on a genome wide scale in premeiotic spermatogonial cells. This allows for the production of a number of genetically unique offspring from a single ENU-mutagenised male to create a variant library.
ENU Mutagenesis can be conducted on an inbred strain or on sensitised background, such as an established transgenic/knockout line.
As ENU-derived dominant variants are identified at the first generation, researchers are able to undertake proof-of-principle evaluations on the variant strains in short timeframes. Many of the variant mice will contain point mutations, meaning that they are more reflective of the mutations that occur in the human genome.
The APN Dominant ENU service is accessible through The Walter and Eliza Hall Institute, Melbourne, the Queensland Institute of Medical Research, Brisbane, and The Menzies Research Institute, Hobart. Staff at these institutions have extensive training and experience in the care and production of mouse strains.
The Dominant ENU nodes provide access to mice heterozygous for random ENU-induced mutations. Access is open to any not-for-profit researcher who can integrate into the current research program without compromising output for other customers. Priority is allocated on a first come, first served basis. As with all APN activities, promotion for the Dominant ENU service will be coordinated by the APN project officer based at ANU in Canberra, and via the APN website and newsletters. In addition, when mice with defined characteristics are identified, the WEHI node manager (Benjamin Kile) will approach Australian researchers with skills in the relevant in the area.
Funding from federal government through the National Collaborative Research Infrastructure Strategy (NCRIS) and State funding from the Victorian Government allows researchers and other collaborators to access Dominant ENU generated mice at a reduced cost.
What is included in the service:
- Creation of ENU-dominant mouse strains
Nodes involved
The Walter and Eliza Hall Institute
Queensland Institute of Medical Research
The Menzies Research Institute
The Centenary Institute of Cancer Medicine and Cell Biology
Veterinary Services Division of the Institute of Medicine and Veterinary Sciences (IMVS) Adelaide
Contact:
T: +61 3 9345 2510
Dr Benjamin Kile
F: +61 3 9347 0852
WEHI Node Manager
The Walter and Eliza Hall Institute
1G Royal Parade
PARKVILLE VIC 3050
Pathology
Histopathology and Organ Pathology
Histopathology and Organ Pathology
Overview
The Histopathology and Organ Pathology service helps researchers across Australia in histological preparation and in analysing histological images and data on specific mice or mice at certain developmental stages. It also generates histological and organ pathology data on mouse models created through the APN.
This service has Digital Slide Scanners (see Zeiss Mirax Digital Slide Scanner details below) capable of producing high quality digitally accessed images of stained slides. Data files are downloadable by e-research fibre-optic links through the IMVS. It also offers the latest Histology capabilities, including confocal microscopes, multi-colour fluorescence microscopy and high resolution white-light microscopy all with electronic image capture.
Staff providing this service have extensive histology, diagnostic and electronic imaging experience. A comparative Pathologist is available at the IMVS site and the Melbourne site has secured the services of consultant Medical and Veterinary Pathologists to provide expert advice.
The APN Histopathology and Organ Pathology service is based at the Department of Anatomy and Cell Biology at The University of Melbourne and through the Veterinary Service Division at the Institute of Medical and Veterinary Science (IMVS) in Adelaide. Samples can be sent to either location.
Zeiss Mirax Digital Slide Scanner
The Zeiss Mirax Digital Slide Scanner is a high resolution, high through-put scanning device capable of imaging standard stained histological slides and slides with specialist staining. Digital images are saved to the server and can be interrogated by the client using the free Mirax Teleconsulting software www.zeiss.com/mirax as required. This service is due to commence in May 2008 and updates on the service will be posted on the Department of Anatomy and Cell Biology web page www.anatomy.unimelb.edu.au under Digital Slide Scanner Services.For more information contact Tina Cardamone at The University of Melbourne, Department of Anatomy and Cell Biology: t.cardamone@unimelb.edu.au
Cost structure
To be determined
Standard Operating Procedure (SOP)
Histopathology and organ pathology SOP documents
Forms
Request for investigation by APN Histopathology and Organ Pathology Service
Information for transportation of GM mice
Nodes involved
Anatomy and Cell Biology at The University of Melbourne
Veterinary Services Division of the Institute of Medicine and Veterinary Sciences (IMVS) Adelaide
Clinical Pathology
The Australian Phenomics Network can now provide a service of haematological analysis of mouse blood samples.
An Advia 2120 haematology analyser is available at the Australian Phenomics Facility in Canberra.
This machine is capable of analysing;
- Complete Blood Counts
- Differentials
- Reticulocytes
The machine has the capability of running mouse samples using its multispecies software package and currently has reference ranges for C57Bl/6 mice and Balb mice.
- The machine requires a minimal sample volume of 175uL to perform a screen. We are also currently developing a dilution system, whereby smaller volumes of blood may be able to be used.
- We ask for a minimum of 200uL to be shipped to the facility at this stage.
The Australian Phenomics Facility is available to run samples for Australian researchers on a cost recovery basis.
- Blood samples can be shipped to the facility for analysis. Samples should preferably arrive Tuesday to Thursday to ensure prompt sampling. A minimum of 48 hours notice is required prior to shipment.
Sampling
Blood is to be collected in EDTA. The method of collection is at the discretion of the researcher, however, it is important to note that the sample should be collected into EDTA as soon as possible to minimise clot formation. Ethics approval may be required from your facility for samples taken from live mice.
- Samples must be of excellent quality with no clots as this will give erroneous results and potentially inhibit the ability to run the sample.
Shipping
- Samples should be shipped on frozen gel packs in an esky to maintain a cool temperature (below 4°C).
For further information on cost and how to collect, ship and organise analysis of blood samples please contact:
Veterinary Services (ANU Bioscience Services)
T: (02) 6125 7755
David Way is the technician in charge of the daily maintenance and running of the machine.
Contact: david.way@anu.edu.au
Dr Suzanne Fowler is the veterinarian in charge of the equipment and procedures.
Contact: suzanne.fowler@anu.edu.au (On leave June 2008 – December 2008).
Node involved
The Australian Phenomics Facility at The Australian National University
Contact:
T: +61 3 8344 4782
Tina Cardamone
F: +61 3 9347 9619
The University of Melbourne Node Manager
Histopathology Service
Department of Anatomy and Cell Biology
Grattan Street
PARKVILLE VIC 3010
Strain Repository
The Strain Repository service comprises of two components:
Both services are provided through the Australian Phenome Bank (APB).
Strain Archiving Service
The APB archives mouse strains as cryopreserved sperm or embryos. The aim of this service is 3-fold:
- it acts as an insurance against disaster;
- archiving removes the cost and effort of maintaining strains “on the shelf” just in case they are needed in the future and;
- archiving preserves strains for future use.
The depositing researcher maintains ownership of the strain and can nominate/confirm availability to other researchers.
Financial support for sperm/embryo cryopreservation is available if the strain:
- has not been frozen in the APB previously;
- has a heritable phenotype, and;
- it will be made available to other researchers, immediately if already in public
domain (published) or after a maximum period of 2 years if not currently in
the public domain.
Mouse Strain Database
The APB maintains, with researcher input, a database held around Australia of live and frozen stock. The database allows researchers to search and determine availability of strains of interest to their work. This contributes to saving research dollars by avoiding multiple importations or duplication of effort to produce strains that are already available in the country as either live or frozen stock.
As strains are created or imported by researchers, they are able to register the strain within the APB. The listing researcher maintains ownership of the strain and can nominate the availability to other researchers, including passing on existing terms and conditions of access.
The database lists fields such as phenotype, genotype, affected gene (if known), allele (if known), method of production (e.g. Tg/KO/ENU) and status (frozen or alive). The database also links to other databases such as Mouse Genome Informatics (MGI) and PubMed.
Benefits to you ...
As a requesting investigator:
- Access to unique models
- Access to existing strains of use to your research
As a donating investigator:
- Fulfills your obligation to share publicly funded and published biomedical research resources
- Frees up animal housing resources
- Reduces per diem costs
- A cryopreserved archive of the line is created
- Eliminates the shipment of mice to multiple requesting investigators
Please use the link to the Phenome Bank on the left or click here to access the database and further information.
Nodes involved
The Australian Phenomics Facility at The Australian National University
Monash Institute of Medical Research at Monash University
Animal Resources Centre (ARC)
Contact:
T: +61 2 6125 1324
Dr Stuart Read
F: +61 2 6125 1381
Phenome Bank Curator
Australian Phenomics Facility
Hugh Ennor Building
Garran Road
ACTON ACT 0200
Further info
Depositor is eligible for extended embryo support if:
- All of the basic support criteria above are met and,
- Strain cannot be frozen down as sperm due to loss of genetic integrity e.g. congenic strains.
Support does not cover:
- Production of donors, depositor is responsible for the cost of producing the required number of animals of correct sex and age,
- Transportation of animals to cryopreservation facility,
- Strain quality control.
Costs to researcher*
|
Sperm |
Embryo |
If eligible for basic subsidy |
$0 |
POA |
If eligible for extended subsidy |
N/A |
POA |
Caging at cryo facility |
$0 |
Caging requirements are strain dependant |
Quality control |
Subjective microscopy analysis test thaw - $60/10 mice |
$450/strain Test thaw and incubation to blastocysts stage - $165/strain |
*Prices are subject to variation without notice.