About

Genetically Engineered Murine Models (GEMM) Core

The Genetically Engineered Murine Models (GEMM) Core provides full-service mouse genetic engineering support to researchers.  We will help you select the best model for your research program, design and build any requisite constructs and reagents, and produce live mice.  The most popular requests are for CRISPR-based gene edited mice and transgenic mice.  To date, we have created engineered alleles for knockouts, conditional knockouts, SNP knockins, tag and marker insertions, deletions of up to 300kb, duplications, and transgenic alleles. These novel mouse models have been made on a wide spectrum of genetic background strains, including but not limited to C57BL/6J, C57BL/6N, NOD, BALB/cJ, FVB/Crl, NSGS, various 129 substrains, collaborative cross strains, and on mice that are already genetically modified.  

Our robust breeding and genotyping program supports breeding to establish germline transmission and can also create cohorts for investigators.  We also provide strain cryopreservation, rederivation and rescue services, as well as cryostorage.  Our services are available to outside institutions, nationally and internationally.

Overview of Services

Gene editing services

  1. Consultation- meet to discuss project goals
  2. gRNA design/construction- with information provided by the investigator, GEMM will design/generate CRISPR reagents for the production of mouse models or for investigator led efforts to modify cell lines
  • Simple- includes design of gRNA to drive a single dsDNA break for the introduction of SNPs, small tag knockins
  • Complex- includes the design of gRNA for dsDNA breaks at two locations in the genome, as necessary for conditional gene knockouts
  1. gRNA activity testing- GEMM will test guide activity in vitro or in embryos cultured to blastocysts
  2. HDR template design and synthesis- GEMM will design HDR templates, synthesize and prepare them for use. This may include the generation of AAV to deliver the desired HDR template.
  3. Microinjection or electroporation- GEMM will generate embryos and introduce gene editing reagents, perform embryo transfers
  4. See breeding program services for genotyping services to identify putative founders

 

Transgenic mouse services

  1. Consultation- meet to discuss project goals. If a transgene has already been created, please complete a service form to initiate the project and skip to step ‘3’.
  2. Transgenic vector design and construction- GEMM will propose a strategy to generate the desired construct for researcher review and approval prior to project start. Once design is approved, GEMM will create the DNA constructs/
  3. Vector preparation for microinjection
  4. Pronuclear injection- GEMM will generate embryos, microinject, and perform embryo transfers
  5. See breeding program services for genotyping services to identify putative founders

 

 

Assisted Reproductive technologies services

  1. Sperm cryopreservation- GEMM will isolate and cryopreserve sperm from up to three individual male mice provided by the researcher
  2. Hybrid cryopreservation- GEMM will isolate and cryopreserve sperm from up to three individual male mice provided by the researcher and in parallel will perform IVF and cryopreserve resulting embryos
  3. Rederivation- using cryopreserved embryos or generating embryos by IVF using fresh or cryopreserved sperm, GEMM will perform embryo transfers to generate live mice
  4. Cryostorage of samples- we provide long term storage of cryopreserved samples, charging an annual fee for this service

 

Breeding program services

  1. Colony management- GEMM will charge a set per diem rate for mice in the breeding program; this per diem charge includes routine handling of the mice to set up mating pairs, check for plugs, and weaning of animals
  2. Genotyping by standard PCR- GEMM will extract gDNA and genotype samples by standard PCR and will perform restriction enzyme digest and/or Sanger sequencing on these PCR products as appropriate
  3. Next generation sequencing support- GEMM will work with the researcher to develop, implement, and analyze results from NGS to genotype mice

 

Other GEMM core services

  1. Blastocyst injection- we will help investigators inject their ES cells into blastocysts upon request
  2. Maxiprep- high copy plasmids, irrespective of eventual use, can be isolated by GEMM for your research
  3. Consumables/supplies- if your project requires additional consumables not accounted for in the established core facility pricing, we will adjust for this. For instance, if you want us to use mice for IVF that are not B6 or NOD, we can adjust for the difference in the cost to purchase, ship and superovulate these animals by charging for a difference in consumables pricing for the service.
  4. Core/labor- GEMM Core personnel’s time will be charged for additional services provided


Location and hours of operation

Hours/Days Open     
Monday-Friday    
8am-5pm    
weekends for animal care    

Links and Resources

https://researchcores.cuanschutz.edu/

https://www.nationaljewish.org/research-science/core-laboratories/mouse-genetics

IACUC : https://research.cuanschutz.edu/committee-support/home/iacuc

Mouse Genome Informatics website: https://www.informatics.jax.org/

Contacts

Name Role Phone Email Location
Jennifer Matsuda

 

 
jennifer.matsuda@cuanschutz.edu