Transgenic/Knockout Mouse Core Facility - Norris Comprehensive Cancer Center - University of Southern California

Summary

The USC/Norris Comprehensive Cancer Center Transgenic Core Facility. The Facility was established in 1993 to provide high-quality embryo and stem cell manipulation, microinjection and related services to the USC community, as well as outside researchers. The core handles the production of transgenic mice, gene targeting and the production of knockout Mice, Construction targeting vector design, preparation of DNA for injection and In vitro fertilization, Re-derivation, as well as Cryopreservation. The Facility is located in both Norris Cancer Center and Zilkha Neurogenetic Institute of USC Medical Center. It comprises a laboratory for DNA purification and quantitation; a cell culture room for performance of ES cell culture, electroporation, pick up clones; a microinjection room for isolation, microinjection and implantation of the fertilized mouse embryos, as well as adjoining vivaria.

 

Equipment/Services

Pronuclear Injection Transgenic Mouse
ES Cell Injection "Knock-out" Mouse

ES Cell Gene Targeting

• Advice on vector design.
• Electroporation (TG/KO provides ES cells).
• Selection of 200-400 ES cell colonies.
• Screening of ES cells for homologous recombinants.
• Additional services available for additional charges.
• Customer performs by PCR or Southern blot.
• Injection of ES cells into blastocysts, please see our "Knock-Out" order form

In Vitro Fertilization

• Reconstitute live mice from fresh or frozen sperm.
• Provides a variety of options for recovering mouse strains endangered by poor breeding, health issues or other problems.
• To generate a large cohort of same-aged mice in one step.

Brief Procedure:

The P.I. provides one or more males (females are included if special strain required) and the Core Facility will harvest oocytes from up to 8 superovulated females and fertilize them in vitro with sperm harvested from the male(s). All 2-cell stage cultured embryos will be implanted into pseudopregnant foster mothers. There are no guarantees regarding the number of pups produced, but if no pups are obtained, the Core Facility will repeat the procedure once at no extra charge.

Additional Services for ES cells

Re-derivation by Embryo Transfer

Embryo Rederivation is used to:

1.  Create a pathogen free strain of mice from lines that are contaminated with various pathogens (MHV, Pinworm, Parvovirus).
2.  Thaw frozen embryos and bring them back to life.

Brief Procedure:
For mice infected with pathogens, the P.I. coordinates the receipt of at least 4 males under the P.I.'s animal care program number.  The Core acquires Wt B6D2F1 females (P.I. must provide the females if a specific background is required) and schedules the procedure for cleaning up the line via embryo transfer.  For frozen embryos, the P.I. transfers embryos in liquid nitrogen and the Core thaws the embryos, and performs implantation.

	# mice prepared
Hybrid or mixed strain	4 or 5 males
Inbred or special strain	5 males and 10 females

Mouse Embryo Cryopreservation

The storage of mouse embryos in liquid nitrogen offers a safe way to preserve lines with potential future use but no current use.  This also saves facility space and the expense associated with keeping a breeding colony.  Another advantage is that during the process of embryo collection, most pathogenic organisms are excluded.  For cryopreservation of a mouse line, the P.I. coordinates the receipt of at least 10 males ages 2.5 - 6 months of the desired strain under the P.I.'s animal care program number.  The Core acquires Wt B6D2F1 females (P.I. must provide the females if a specific background is required) and schedules the freezing 200 or more fertilized eggs from each line in straws.  As a quality control, the Core will thaw a straw of embryos along with each frozen line and will culture them to the blastocyst stage.

	# mice prepared
Hybrid or mixed strain	8 males or more
Inbred or special strain	10 males and 20 females

Special Services

Price List

Click here for pricing information.

Turn Around Time

Transgenic Mouse Orders:
We will pay for the maintenance of foster mothers and pups up to three weeks from date of birth. During that time we expect the PI to

1. Genotype the mice
2. Transfer the mice to the PI's vivarium protocol. We will charge double for the mice left on the Core protocol after three weeks. In general, the injection will be repeated until at least 2 transgenic pups are obtained (except the cases in which the transgenic causes embryonic lethality). Transgenic status refers only to DNA integration as assesed by Southern blot. We can't guarantee expression of an RNA or protein product from the integrated DNA.

Abstract

Please contact us for more information.

More information

Website

Visit the Website

Location

USC/Norris Comprehensive Cancer Center Transgenic Core Facility
1441 Eastlake Ave. Room 5318
Los Angeles, CA 90089-9176
Telephone: (323) 865-0726
Fax: 323/865-0098
Email: CoreTransgenic@ccnt.usc.edu

Contact Information

Robert Maxson
Director
Telephone: (323) 865-0633
Fax: (323) 865-0098
Email: maxson@usc.edu

Nancy Lanying Wu, MD
Telephone: (323) 865-0726
Fax (323) 865-0098
Email: nwu@usc.edu

Youzhen Yan Injection
Technician
Telephone: (323) 865-0639
Fax: (323) 865-0098
Email: youzhen@usc.edu

John Johnson
Financial Assistant
Telephone: (323) 865-0639
Fax: (323) 865-0098
Email: john.e.johnson@usc.edu

Linda Liu, PhD
Gene Targeting Manager
Telephone: (323) 865-0635
Fax: (323) 865-0098
Email: liqiongl@usc.edu

Scheduling Appointments

Transgenic/Knockout Services:
Nancy Lanying Wu, MD
Manager
Telephone: (323) 865-0726
Email: nwu@usc.edu

Gene Targeting Services: 
Mandy Man-Chun Ting, PhD
Manager
Telephone: (323) 865-0615
Email: manchunt@usc.edu