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The Transgenic Gene-Targeting Core has been an integral part of the Gladstone Institutes UCSF research community for more than 20 years, producing essential mouse models as a service to Gladstone investigators and UCSF faculty.
To generate a transgenic mouse, a fragment of DNA is injected through a glass needle into a pronucleus of a fertilized one-cell host embryo. Microinjected embryos are implanted into oviducts of recipient female mice to permit development of the embryos into new lines of “transgenic” animals with the foreign DNA incorporated into their genomes. The foreign DNA can be designed to contain a functional gene that produces a particular protein or modulates the expression of host genes.
To generate a knockout mouse, a targeting construct is electroporated into ES cells, colonies are selected for the desired mutation. The ES cell clones are injected into the blastacoel cavity of a day 3.5 embryo. Microinjected embryos are implanted into the uterus of recipient female mice to permit development of the embryos into chimeras, mice that contain a mix of host and injected cells. The chimeras are then bred to produce heterozygote mice that are then crossed to produce a knockout mouse line.
CRISPR/Cas9 knock-out (KO) and knock-in (KI) technology efficiently creates gene knock-out (KO) and knock-in (KI) mice. This new technology can also be used to generate mice carrying mutations in multiple genes in one step . The core provides services for making CRISPR KO and KI mice. Cas9, sgRNA, and donor DNA can be injected or electroporated on C57BL/6, FVB and other strains. Contact the core manager if you wish to use another strain.
Transgenic and knockout technologies enable investigators to learn how specific genes and proteins, or their variants, function in a natural living animal environment with a defined genetic background in ways not possible by other experimental approaches. At the Gladstone Institutes transgenic mouse models have been used to understand how specific genes affect heart disease, atherosclerosis, Alzheimer's disease, obesity, AIDS, and other diseases and disorders that impact modern life.
The Transgenic Gene Targeting Core creates genetically modified mouse models. The services provided by the core are on a fee-for-service basis for Investigators at the Gladstone Institutes, UCSF, and other research institutions.
Services include:
Junli Zhang - Core Director (primary contact)
Email: junli.zhang@gladstone.ucsf.edu
Phone: 415-734-4019
Fax: 415-355-0960
Location |
Hours |
The J. David Gladstone Institutes 1650 Owens Street, San Fransisco, CA 94158
|
9am - 5pm, Monday - Friday
|
Name | Role | Phone | Location | |
---|---|---|---|---|
Junli Zhang |
Research Scientist, Core Director
|
415-734-4019
|
junli.zhang@gladstone.ucsf.edu
|
|
Isidro Espineda |
Sr. Research Technologist
|
415-734-4002
|
sid.espineda@gladstone.ucsf.edu
|
|
Carlos Benitez |
Research Associate
|
415-734-4040
|
carlos.benitez@gladstone.ucsf.edu
|
|
Carlisa Benitez |
Lab Associate
|
carlisa.benitez@gladstone.ucsf.edu
|