Mouse Genetics
The Mouse Genetics Core Facility provides investigators the possibility of generating genetically engineered mice by transgenic and gene targeting methodologies. The core can generate, genetically manipulate, and select mouse embryonic stem (ES) cells from various genetic strains for injection into blastocysts. Alternatively, these genetically modified ES cells can be differentiated in vitro by the core facility into various cell types for investigator research. The core also offers injection of transgenes into fertilized oocytes.
The facility can advise investigators on the generation of transgenic and targeting constructs for simple knock-out/knock-in and for tissue-specific and conditional mutations as well as strategies for screening transfected ES cells and mouse mutants. In addition, the core facility performs rederivation of "dirty" mice by embryo transfer and cryopreservation of mouse strains (embryos).
Our Services
The following is a list of services commonly provided by our facility. Please direct inquiries about these services or any additional services not listed below by emailing us at MouseGenetics@NJHealth.org. We can readily provide you with forms, timelines, and prices for any given service. We also have an extensive collection of references and protocols compiled for your use.
Design and Generation of Constructs
We will help you develop and design constructs for your
research. We make constructs for BAC transgenics, conventional
transgenics, and gene targeting. We can also make expression constructs
for your research. Learn more.
Gene Targeting in Mouse ES Cells
Targeting mouse embryonic stem cells for a desired genetic modification. Learn more.
ES Cell Screening for Homologous Recombinants
Loss of Allele, a quantitative PCR technique, will be used to rapidly identify putative positives for secondary screening.
This service includes the design, purchase, and validation of suitably positioned primer/probe pairs for your targeting construct, preparation of genomic DNA from targeted ES cell clones, and loss of allele (LOA) analysis. If the primer/probe set passes our validation tests (for primer efficiency), the screening of ES clones can be quite rapid, within a month or less.
Blastocyst Injection
Production of chimeric mice by injecting targeted ES cells into blastocyst stage embryos. Learn more.
Pronuclear Injection
Production of potential transgenic founder mice by injecting a DNA construct into the pronucleus of zygotes. Learn more.
Cryopreservation of Mouse Embryos
Cryopreservation and long-term storage of 8-cell stage mouse embryos in liquid nitrogen. Learn more.
Cryopreservation of Mouse Sperm
Sperm will be
cryopreserved and placed in long term storage in liquid nitrogen. We
will verify the quality of frozen sperm by IVF.
Sperm from 2 to 3 males of the desired genotype will be harvested and cryopreserved from 14-20 week old proven males, which have been actively bred and then rested for 5-7 days before sperm harvest. Twenty straws of sperm will be cryopreserved. The quality of the cryopreserved sperm will be verified by IVF with wildtype C57Bl/6 eggs. In addition, annual cryostorage fees will be charged to the investigator based upon the number of straws.
Speed Cryopreservation
Sperm will be harvested from
one or two proven male mice to generate 2-cell stage embryos, which will
be stored in liquid nitrogen.
Twenty-five C57Bl/6 females will be purchased for your project, superovulated, and unfertilized eggs will be collected. Fresh sperm will be harvested from 1 or 2 males of the desired genotype that have proven breeding success and will be used with the eggs for in vitro fertilization. All healthy 2-cell stage embryos will be cryopreserved for the investigator. If 300 or more 2-cell stage embryos are cryopreserved, a test rederivation will be performed, transferring embryos into pseudopregnant females for the generation of live pups. In addition, annual cryostorage fees will be charged to the investigator based upon the number of straws of embryos stored ($50 for 1-25 straws/vials).
Rederivation of Mouse Lines
Rederivation of mouse lines from cryopreserved embryos. Learn more.
Rederivation of Mouse Lines by IVF
We can rederive
mouse lines from fresh or cryopreserved sperm using IVF. Resulting
embryos are transferred into pseudopregnant females to give rise to live
offspring.
This service involves harvesting sperm from a proven male stud(s), ideally ranging from 14-20 weeks of age, (or using cryopreserved sperm) and culturing it with C57BL/6 unfertilized eggs from 10 donor females under IVF conditions to generate embryos. Resulting embryos are transferred into pseudopregnant females.
Chromosome Counting
Determination of chromosome number in ES cell clones. Learn more.
Mycoplasma Testing
Testing of tissue culture supernatant for the presence of mycoplasma. Learn more.
