- routine stock-keeping and maintenance of fly repository, using three stock-keeping robots
- transgenic services, including outsourced injections of P element and Integrase constructs and screening of injection crosses
- mandatory quarantine of all incoming stocks
- assistance with mutant screens
- virgin collection of commonly used stocks
- setting up crosses
- performance of recombination crosses and stock constructions
- stock-keeping: the Fly Facility maintains approximately 22,000 stocks using three stock-keeping robots. Stocks are kept in duplicate, on a four-week cycle, at 18 degrees C, with vials being a week apart in age. Access to the stocks is restricted to Fly Facility personnel only. Stocks are normally delivered within a few hours.
- fly transgenic services: the Facility outsources all transgenic work to Genetic Services, Inc (GSI) in Cambridge, Mass. As the single point of contact between Janelia and GSI, the Facility sends out the DNA, receives the transformants back, and handles all correspondence. The majority of constructs sent out use the Phi C31 integrase system. Constructs are injected and screened for transformants by GSI, and the transformants are run through a series of crosses to remove the integrase and produce homozygous lines. This pipeline feeds the Fly Light and Fly Olympiad projects as well as the individual labs. The Facility processes about 500 transformant vials every month.
General Fly Help
Fly Facility personnel are available to assist with general screening, setting up Gal4 – UAS crosses, and performing recombinations or stock constructions.
- three stock-changing robots
- Jane and Cornelia, Oct 2006
- Pickens, May 2009
- maximum capacity = 24,000 stocks
- manufactured by Keller Swartwood Engineering (KSE)
- originally designed for Exelixis to prevent work place injury
- robots run from 7:00am – 7:00pm Monday - Friday
- one robot can change about 100 stocks/hour
- three robots change 2,700-3,600 stocks/day
- robots send an email when there is a problem
- currently maintain 22,000 stocks
- special features: robots make copies of the stock without changing database; suspend checks, adding data to the database; A to B transfer, adding second copy into the databa
Combined with the services of microinjection, rederivation, and cryopreservation, staff deliver the full package for creation, propagation, and preservation of genetically-engineered animals. The Facility will continue to be the best in production and efficiency.
- Design and generate gene targeting constructs, including knockin report genes, knockin mutations, conventional/conditional knockouts, and BAC transgenics
- ES cell targeting
- Generation of chimeras through ES cell/embryo aggregation
- DNA pronuclear microinjection, including full-service DNA purification, microinjection and housing potential transgenic pups until 3 wks old. At least 3 transgenic mice or rats are guaranteed
- Establish new ES cell lines from blastocysts. The service is provided for derivation of new ES cell lines from wildtype or mutant mouse lines. At least 2 ES cell lines from one mouse line derived are guaranteed. Three vials of ES cells with passage number lower than P7 are provided for each ES cell line.
- Rederivation of contaminated mouse lines or importation of mouse lines by embryo transfer
- Mouse Embryo/sperm Cryopreservation
- Developing new technologies
One of the Facility's missions is to develop and experiment with new technologies to support scientists' research needs.
The GTTF provides consultation related to gene targeting and transgenics. Researchers are welcome to discuss various topics such as transgenic/targeting constructs, vectors for making constructs, ES cell mediated gene targeting, conditional genome alteration, breeding strategy, genotyping analysis and transgenic colony management.
As an AAALAC facility with international accreditation since 2008, the Vivarium uses the latest technologies in research animal housing and care. Our facilities and procedures follow guidelines outlined in the Guide for the Care and Use of Laboratory Animals, published by the National Research Council (NRC); PHS Policy on Humane Care and Use of Laboratory Animals, published by the United States Public Health Service; other applicable federal and state laws and regulations; and best practices in the industry.
Following policies advocated by both the NRC and PHS publications, Janelia maintains an Institutional Animal Care and Use Committee (IACUC). The Senior Director of Scientific Services serves as the Institutional Official who is responsible for all IACUC activities. The IACUC evaluates proposed uses of animals in research, testing, or education; inspects facilities where animals are used in research; evaluates the use of animals in research at Janelia; submits reports; and receives and reviews concerns regarding the use of animals at Janelia.
To the extent possible, the Facility follows the "3R's" put forth by Russell and Burch (1959): reduction in the number of animals used; refinement of techniques to minimize pain or distress; and, when practical, replacement with non-animal techniques and models. In an environment designed to maintain biosecurity and ensure healthy animals, the Facility uses state-of-the-art equipment and procedures, including sterile microenvironment containment cages, robotic cage wash, automated drinking water distribution, and automated water treatment for aquatic species.
Animal Care: Vivarium staff provide the husbandry care for all vertebrate species and some invertebrate species used in research at Janelia. This includes providing food and water, daily health assessments, and cage changes. Species supported include mice, rats, salamanders, dragonfly larvae, and zebrafish.
Breeding: Vivarium staff manage all researcher breeding colonies and provide the animals needed for experiments. This includes setting up matings, weaning, identification, tissue sampling for genetic analysis, and recording information in an electronic database.
Technical Services: Vivarium staff provide assistance with various technical services including food and water restriction support, injections, blood sampling, and necropsies.
Animal Ordering: Vivarium staff order animals on behalf of investigators from approved vendors (Charles River Laboratories, Jackson Laboratories, MMRRC, and Taconic).
Animal Imports/Exports: Vivarium staff coordinate and manage the shipping and receiving of animals from collaborating institutions, including obtaining health reports. All animals are rederived before entering the main breeding colony rooms.
Surgical Support: Vivarium staff members perform several surgeries for researchers, including in utero electroporation, in utero viral vector injection, postnatal viral vector injection, and intracranial cannulation.